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1.
Fisher, Kimberly Day.
Regulation of acid-stimulated H+ secretion by Pyk2 and MAPK signaling pathways in the outer medullary collecting duct.
Degree: 2012, Wake Forest University
URL: http://hdl.handle.net/10339/37423 
► Acid-secreting intercalated cells of the outer medullary collecting duct respond to changes in systemic pH through regulation of apical H+ transporters. Little is known about…
(more)
▼ Acid-secreting intercalated cells of the outer medullary collecting duct respond to changes in systemic pH through regulation of apical H+ transporters. Little is known about the mechanism by which these cells sense changes in extracellular pH (pHo). Pyk2 is a non-receptor tyrosine kinase activated by auto-phosphorylation at Tyr402 by cell-specific stimuli, including decreased pH and is involved in the regulation of MAPK signaling pathways and transporter activity. Therefore we examined whether the Pyk2 and MAPK up-regulate ATPase mediated proton secretion in response to decreased pH in outer medullary collecting duct cells. Immunoblot analysis of phosphorylated Pyk2 (Tyr402), ERK1/2 (Thr202/Tyr204), and p38 (Thr180/Tyr182) was used to assay protein activation. To examine specificity of kinase activation and its effects, we used Pyk2 siRNA to knockdown Pyk2 expression levels, the Src kinase inhibitor PP 1 to inhibit Src phosphorylation, and the MEK inhibitor U0126 to inhibit ERK1/2 phosphorylation. Furthermore, we used adenoviral-gene transfer of AdCRNK, a dominant-interfering truncated Pyk2 construct, to block Pyk2 phosphorylation at Tyr402 as well as the p38-specific inhibitor, SB203580. The pH-sensitive fluorescent probe BCECF-AM was used to assay H+ transporter activity. The activity of H+ transporters was measured as the rate of intracellular pH recovery after an NH4Cl pre-pulse. We show that Pyk2 is endogenously expressed and activated by acid pH in mouse-derived outer medullary collecting duct (mOMCD1) cells. Incubation of mOMCD1 cells in acid media (pHo 6.7) and reduction in pHi induced by an NH4Cl pre-pulse increased the phosphorylation of Pyk2, ERK1/2 and p38. Consistent with our previous studies, we found that mOMCD1 cells exhibit H+-ATPase and H+,K+-ATPase activity. Pyk2 inhibition by Pyk2 siRNA and PP 1 prevented Pyk2 and ERK1/2 phosphorylation as well as H+-ATPase-mediated recovery. Pyk2 inhibition by AdCRNK prevented Pyk2 and p38 phosphorylation as well as H+,K+-ATPase-mediated pHi recovery in mOMCD1 cells. In addition, inhibition by U0126 prevented acid-induced ERK1/2 phosphorylation and H+-ATPase-mediated pHi recovery but not phosphorylation of p38. We conclude that Pyk2 is required for ERK1/2 and p38 signaling pathways that stimulate H+-ATPase and H+,K+-ATPase activity, respectively, in response to acute acidosis in mOMCD1 cells.
Subjects/Keywords: ATPase
…ATPase mediated proton secretion in response
to decreased pH in outer medullary collecting duct… …p38. Consistent with our previous studies, we found that mOMCD1 cells exhibit H+ATPase and H… …K+-ATPase activity. Pyk2 inhibition by Pyk2 siRNA and PP 1 prevented Pyk2
and ERK1/2… …phosphorylation as well as H+-ATPase-mediated recovery.
Pyk2 inhibition by
AdCRNK prevented Pyk2 and… …p38 phosphorylation as well as H+,K+-ATPase-mediated pHi
viii
recovery in mOMCD1 cells…
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APA (6th Edition):
Fisher, K. D. (2012). Regulation of acid-stimulated H+ secretion by Pyk2 and MAPK signaling pathways in the outer medullary collecting duct. (Thesis). Wake Forest University. Retrieved from http://hdl.handle.net/10339/37423
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Chicago Manual of Style (16th Edition):
Fisher, Kimberly Day. “Regulation of acid-stimulated H+ secretion by Pyk2 and MAPK signaling pathways in the outer medullary collecting duct.” 2012. Thesis, Wake Forest University. Accessed April 17, 2021.
http://hdl.handle.net/10339/37423.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Fisher, Kimberly Day. “Regulation of acid-stimulated H+ secretion by Pyk2 and MAPK signaling pathways in the outer medullary collecting duct.” 2012. Web. 17 Apr 2021.
Vancouver:
Fisher KD. Regulation of acid-stimulated H+ secretion by Pyk2 and MAPK signaling pathways in the outer medullary collecting duct. [Internet] [Thesis]. Wake Forest University; 2012. [cited 2021 Apr 17].
Available from: http://hdl.handle.net/10339/37423.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Fisher KD. Regulation of acid-stimulated H+ secretion by Pyk2 and MAPK signaling pathways in the outer medullary collecting duct. [Thesis]. Wake Forest University; 2012. Available from: http://hdl.handle.net/10339/37423
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
University of New Mexico
2.
Chan, Chun-Yuan.
Yeast V-ATPase Regulation by Phosphofructokinase-1.
Degree: Biomedical Sciences Graduate Program, 2015, University of New Mexico
URL: https://digitalrepository.unm.edu/biom_etds/118
► V-ATPase is a vacuolar (lysosome-like) ATPase-dependent proton pump necessary for maintaining pH homeostasis in the organelles of the endomembrane system. It also contributes to regulation…
(more)
▼ V-
ATPase is a vacuolar (lysosome-like)
ATPase-dependent proton pump necessary for maintaining pH homeostasis in the organelles of the endomembrane system. It also contributes to regulation of the cytosol pH and the extracellular pH. In specialized cells (renal intercalated cells, epididymis clear cells, and osteoclasts), V-
ATPase proton transport supports urinary acidification, sperm maturation, and bone resorption. Genetic mutations of V-
ATPase expressed in those tissue-specific cells cause distal renal tubular acidosis, infertility, and osteopetrosis.
V-ATPases are composed of a peripheral domain (V1), which hydrolyzes ATP, and a membrane-bound domain (Vo), which transports proton. V-
ATPase activity is tightly regulated in vivo by numbers of mechanisms, including reversible disassembly of the V1 and Vo domains. Glucose, the nutrient oxidized in glycolysis, modulates reversible dissociation of V-
ATPase. This dissertation was aimed at understanding how subunits of phosphofructokinase-1 (α subunit and β subunit) regulate V-
ATPase function. Our results showed that both subunits are important for V-
ATPase activity, but β subunit displayed more significant phenotypes. Deletion of β subunit reduced glucose-dependent V1Vo reassembly and altered V-
ATPase binding to its assembly factor, RAVE. We additionally investigated the mechanisms by which phosphofructokinase-1 controls V-
ATPase function. We concluded that glucose-dependent V1Vo reassembly and V-
ATPase function at steady state were controlled by the glycolytic flux, independently of phosphofructokinase-1. Notably, V-
ATPase activation in vivo correlated with the presence of phosphoglycerate kinase at vacuolar membranes. These studies further advanced our understanding how glucose controls V-
ATPase pumps in vivo.
Advisors/Committee Members: Parra, Karlett J., Osley, Mary Ann, Garver, William S., Lee, Samuel A..
Subjects/Keywords: V-ATPase
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APA (6th Edition):
Chan, C. (2015). Yeast V-ATPase Regulation by Phosphofructokinase-1. (Doctoral Dissertation). University of New Mexico. Retrieved from https://digitalrepository.unm.edu/biom_etds/118
Chicago Manual of Style (16th Edition):
Chan, Chun-Yuan. “Yeast V-ATPase Regulation by Phosphofructokinase-1.” 2015. Doctoral Dissertation, University of New Mexico. Accessed April 17, 2021.
https://digitalrepository.unm.edu/biom_etds/118.
MLA Handbook (7th Edition):
Chan, Chun-Yuan. “Yeast V-ATPase Regulation by Phosphofructokinase-1.” 2015. Web. 17 Apr 2021.
Vancouver:
Chan C. Yeast V-ATPase Regulation by Phosphofructokinase-1. [Internet] [Doctoral dissertation]. University of New Mexico; 2015. [cited 2021 Apr 17].
Available from: https://digitalrepository.unm.edu/biom_etds/118.
Council of Science Editors:
Chan C. Yeast V-ATPase Regulation by Phosphofructokinase-1. [Doctoral Dissertation]. University of New Mexico; 2015. Available from: https://digitalrepository.unm.edu/biom_etds/118
University of Alberta
3.
Horvat, Natalie K.
Characterization of the motif requirements for the function
of Aha1p and its homologue Hch1p.
Degree: MS, Department of Cell Biology, 2013, University of Alberta
URL: https://era.library.ualberta.ca/files/2r36tz13q
► The Hsp90 chaperone facilitates the maturation of client proteins, which are key players in cancer. Hsp90 inhibitor drugs, such as NVP-AUY922, are promising anti-cancer therapies.…
(more)
▼ The Hsp90 chaperone facilitates the maturation of
client proteins, which are key players in cancer. Hsp90 inhibitor
drugs, such as NVP-AUY922, are promising anti-cancer therapies.
Co-chaperones regulate the ATPase-dependent Hsp90 activity and
specifically, the co-chaperone Aha1 is the most robust stimulator
of Hsp90 ATPase activity. Hch1p is a homologue of Aha1p and shares
numerous conserved motifs. The conserved RKxK motif is involved in
remodeling of the catalytic loop in Hsp90 and is required for Hch1p
and Aha1p function. Surprisingly, the highly conserved N terminal
peptide NxxNWHW is required for Hch1p activity in vivo but not for
ATPase stimulation by either co-chaperone in vitro. Interestingly,
Hch1p regulates sensitivity to Hsp90 inhibitor drugs in vivo
whereas Aha1p does not. I propose that Hch1p regulates a step
distinct than that of Aha1p which occurs early in the Hsp90 cycle
and is sensitive to drug inhibition.
Subjects/Keywords: ATPase; Hch1; Hsp90
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APA ·
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APA (6th Edition):
Horvat, N. K. (2013). Characterization of the motif requirements for the function
of Aha1p and its homologue Hch1p. (Masters Thesis). University of Alberta. Retrieved from https://era.library.ualberta.ca/files/2r36tz13q
Chicago Manual of Style (16th Edition):
Horvat, Natalie K. “Characterization of the motif requirements for the function
of Aha1p and its homologue Hch1p.” 2013. Masters Thesis, University of Alberta. Accessed April 17, 2021.
https://era.library.ualberta.ca/files/2r36tz13q.
MLA Handbook (7th Edition):
Horvat, Natalie K. “Characterization of the motif requirements for the function
of Aha1p and its homologue Hch1p.” 2013. Web. 17 Apr 2021.
Vancouver:
Horvat NK. Characterization of the motif requirements for the function
of Aha1p and its homologue Hch1p. [Internet] [Masters thesis]. University of Alberta; 2013. [cited 2021 Apr 17].
Available from: https://era.library.ualberta.ca/files/2r36tz13q.
Council of Science Editors:
Horvat NK. Characterization of the motif requirements for the function
of Aha1p and its homologue Hch1p. [Masters Thesis]. University of Alberta; 2013. Available from: https://era.library.ualberta.ca/files/2r36tz13q
University of Alberta
4.
Mai, BaoChan N.
The Biochemical Characterization of the ATPase activity of
three Hsp82 point mutants: Hsp82pA587T, Hsp82pG313S, and
Hsp82pE381K.
Degree: MS, Department of Cell Biology, 2012, University of Alberta
URL: https://era.library.ualberta.ca/files/c534fp07w
► Chaperones are family of proteins that assist in protein folding. 90 kDa Heat shock protein (Hsp90 mammalians, Hsp82 in Saccharomyces cerevisiae) is a well conserved…
(more)
▼ Chaperones are family of proteins that assist in
protein folding. 90 kDa Heat shock protein (Hsp90 mammalians, Hsp82
in Saccharomyces cerevisiae) is a well conserved chaperone that is
essential for eukaryotic viability. The Hsp90 cycle is regulated by
the ability to hydrolyze ATP, and through the interactions with
other proteins known as co-chaperones. I biochemically
characterized three Hsp82 point mutants: Hsp82pA587T, Hsp82pG313S,
and Hsp82pE381K, using co-chaperones known to influence the ATPase
activity of Hsp82p (Aha1p, Sti1p, Sba1p, and Hch1p). The ATPase
activity of the Hsp82pG313S mutant could not characterize due to
the low signal to noise ratio. I discovered the Hsp82pA587T mutant
ATPase activity was over stimulated by Aha1p, but had a similar
relationship as the wild-type in terms of the Sti1p and Sba1p. With
the Hsp82pE381K mutant, I observed that the mutant was not
stimulated robustly by Aha1p, and this stimulated rate was not
inhibited by Sti1p.
Subjects/Keywords: Hsp82; ATPase; Biochemical
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APA ·
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Manager
APA (6th Edition):
Mai, B. N. (2012). The Biochemical Characterization of the ATPase activity of
three Hsp82 point mutants: Hsp82pA587T, Hsp82pG313S, and
Hsp82pE381K. (Masters Thesis). University of Alberta. Retrieved from https://era.library.ualberta.ca/files/c534fp07w
Chicago Manual of Style (16th Edition):
Mai, BaoChan N. “The Biochemical Characterization of the ATPase activity of
three Hsp82 point mutants: Hsp82pA587T, Hsp82pG313S, and
Hsp82pE381K.” 2012. Masters Thesis, University of Alberta. Accessed April 17, 2021.
https://era.library.ualberta.ca/files/c534fp07w.
MLA Handbook (7th Edition):
Mai, BaoChan N. “The Biochemical Characterization of the ATPase activity of
three Hsp82 point mutants: Hsp82pA587T, Hsp82pG313S, and
Hsp82pE381K.” 2012. Web. 17 Apr 2021.
Vancouver:
Mai BN. The Biochemical Characterization of the ATPase activity of
three Hsp82 point mutants: Hsp82pA587T, Hsp82pG313S, and
Hsp82pE381K. [Internet] [Masters thesis]. University of Alberta; 2012. [cited 2021 Apr 17].
Available from: https://era.library.ualberta.ca/files/c534fp07w.
Council of Science Editors:
Mai BN. The Biochemical Characterization of the ATPase activity of
three Hsp82 point mutants: Hsp82pA587T, Hsp82pG313S, and
Hsp82pE381K. [Masters Thesis]. University of Alberta; 2012. Available from: https://era.library.ualberta.ca/files/c534fp07w
5.
宮本, 健志.
ラット脳各種ATPase活性に対する静脈麻酔薬の作用 : Effects of intravenous anesthetics on the activities of various ATPases in rat brain.
Degree: 博士(歯学), 2014, Hokkaido University / 北海道大学
URL: http://hdl.handle.net/2115/58182
;
http://dx.doi.org/10.14943/doctoral.k11272
► The effects of general anesthetics on functional proteins remain to be clarified. This study examined whether the activity of magnesium-dependent ATPase (Mg2+ -ATPase) in the…
(more)
▼ The effects of general anesthetics on functional proteins remain to be clarified. This study examined whether the activity of magnesium-dependent ATPase (Mg2+ -ATPase) in the rat brain is a target for intravenous anesthetics. The effects of propofol, pentobarbital, and thiopental on magnesium-dependent ATPase in the plasma membrane fraction (PII) and microsomal fraction (PIII) isolated from rat brain homogenates by the method of Pottorf were examined. The optimal pH values for Mg2+ -ATPase activities in PII and PIII were 9.4 and 7.4, respectively. The Mg2+ -ATPase activity of PII was inhibited by about 80% by NaN3(sodium azide), an inhibitor of F-type ATPase. The Mg2+ -ATPase activity of PIII was inhibited by about 30% by the V-type ATPase inhibitor bafilomycin, and by about 10% by the P-type ATPase inhibitor vanadate. Western blotting analysis revealed that the largest amount of F-type ATPase was detected in PII, and the largest amount of V-type ATPase was detected in PIII. We defined all ATPase activity that was insensitive to the above inhibitors as basal Mg2+ -ATPase activity. In the presence of these inhibitors, we examined the effects of intravenous anesthetics on the activities of various ATPases. Propofol inhibits V-type ATPase activity, PII basal Mg2+ -ATPase activity, and PIII basal Mg2+ -ATPase activity in a dose-dependent manner; however, F-type ATPase activity was uniquely stimulated by about 80% by propofol at concentrations below 0.6 mM. Pentobarbital inhibited all types of ATPases in a dose-dependent manner. Thiopental inhibited all types of ATPases to varying degrees.
静脈麻酔薬の各種ATPaseに対する作用の報告は多いが，作用機構の詳細については未だ不明な点が多い．ATPaseにはP型ATPase,V型ATPase,F型ATPaseなどが存在し，活性発現にMg2+（Mg）を必要とするが，役割や性質の不明なMg-ATPaseは多数報告されている．本研究ではラット脳に存在するMg-ATPaseを分析するとともに，静脈麻酔薬の作用を検討した．ラット全脳から，Pottorfの方法に従って，形質膜（PII）とミクロソーム（PIII）分画を得た．Mg-ATPase活性の至適pHはPIIが約9.4，PIIIは約7.4であった．PIIのMg-ATPase活性はF型ATPaseの阻害剤であるNaN3によって約80%抑制され，PIIIのMg-ATPase活性はP型ATPase及びV型ATPaseの阻害剤であるNa3VO4及びBafilomycine A1によってそれぞれ約10%及び30%抑制された．ウエスタンブロッティングの結果からも、F型ATPaseはPIIに，V型ATPaseはPIIIに最も多く検出された．上記阻害剤で抑制されないMg-ATPase活性をBasal Mg-ATPaseとし，各阻害剤の存在下で各ATPaseを分別して静脈麻酔薬の影響を調べた．PropofolはV型ATPase活性、PII及びPIII Basal Mg-ATPase活性を濃度依存的に抑制したが，F型ATPase活性は80%程度活性化した．PentobarbitalはすべてのATPase活性を濃度依存的に抑制した．ThiopentalもすべてのATPase活性を抑制したが，その程度は各ATPaseによって異なった．以上の結果から、静脈麻酔薬はラット脳Mg-ATPase活性を基本的に抑制するが，Propofolはミトコンドリアに特異的な作用を及ぼすことが示唆された.
16
Hokkaido University（北海道大学）. 博士(歯学)
Subjects/Keywords: 静脈麻酔薬; F型ATPase; V型 ATPase; Basal Mg2+ -ATPase
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APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
宮本, . (2014). ラット脳各種ATPase活性に対する静脈麻酔薬の作用 : Effects of intravenous anesthetics on the activities of various ATPases in rat brain. (Thesis). Hokkaido University / 北海道大学. Retrieved from http://hdl.handle.net/2115/58182 ; http://dx.doi.org/10.14943/doctoral.k11272
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Chicago Manual of Style (16th Edition):
宮本, 健志. “ラット脳各種ATPase活性に対する静脈麻酔薬の作用 : Effects of intravenous anesthetics on the activities of various ATPases in rat brain.” 2014. Thesis, Hokkaido University / 北海道大学. Accessed April 17, 2021.
http://hdl.handle.net/2115/58182 ; http://dx.doi.org/10.14943/doctoral.k11272.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
宮本, 健志. “ラット脳各種ATPase活性に対する静脈麻酔薬の作用 : Effects of intravenous anesthetics on the activities of various ATPases in rat brain.” 2014. Web. 17 Apr 2021.
Vancouver:
宮本 . ラット脳各種ATPase活性に対する静脈麻酔薬の作用 : Effects of intravenous anesthetics on the activities of various ATPases in rat brain. [Internet] [Thesis]. Hokkaido University / 北海道大学; 2014. [cited 2021 Apr 17].
Available from: http://hdl.handle.net/2115/58182 ; http://dx.doi.org/10.14943/doctoral.k11272.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
宮本 . ラット脳各種ATPase活性に対する静脈麻酔薬の作用 : Effects of intravenous anesthetics on the activities of various ATPases in rat brain. [Thesis]. Hokkaido University / 北海道大学; 2014. Available from: http://hdl.handle.net/2115/58182 ; http://dx.doi.org/10.14943/doctoral.k11272
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
6.
宮本, 健志.
ラット脳各種ATPase活性に対する静脈麻酔薬の作用.
Degree: 博士(歯学), 歯学, 2014, Hokkaido University
URL: http://hdl.handle.net/2115/58182
► The effects of general anesthetics on functional proteins remain to be clarified. This study examined whether the activity of magnesium-dependent ATPase (Mg2+ -ATPase) in the…
(more)
▼ The effects of general anesthetics on functional proteins remain to be clarified. This study examined whether the activity of magnesium-dependent ATPase (Mg2+ -ATPase) in the rat brain is a target for intravenous anesthetics. The effects of propofol, pentobarbital, and thiopental on magnesium-dependent ATPase in the plasma membrane fraction (PII) and microsomal fraction (PIII) isolated from rat brain homogenates by the method of Pottorf were examined. The optimal pH values for Mg2+ -ATPase activities in PII and PIII were 9.4 and 7.4, respectively. The Mg2+ -ATPase activity of PII was inhibited by about 80% by NaN3(sodium azide), an inhibitor of F-type ATPase. The Mg2+ -ATPase activity of PIII was inhibited by about 30% by the V-type ATPase inhibitor bafilomycin, and by about 10% by the P-type ATPase inhibitor vanadate. Western blotting analysis revealed that the largest amount of F-type ATPase was detected in PII, and the largest amount of V-type ATPase was detected in PIII. We defined all ATPase activity that was insensitive to the above inhibitors as basal Mg2+ -ATPase activity. In the presence of these inhibitors, we examined the effects of intravenous anesthetics on the activities of various ATPases. Propofol inhibits V-type ATPase activity, PII basal Mg2+ -ATPase activity, and PIII basal Mg2+ -ATPase activity in a dose-dependent manner; however, F-type ATPase activity was uniquely stimulated by about 80% by propofol at concentrations below 0.6 mM. Pentobarbital inhibited all types of ATPases in a dose-dependent manner. Thiopental inhibited all types of ATPases to varying degrees.
静脈麻酔薬の各種ATPaseに対する作用の報告は多いが，作用機構の詳細については未だ不明な点が多い．ATPaseにはP型ATPase,V型ATPase,F型ATPaseなどが存在し，活性発現にMg2+（Mg）を必要とするが，役割や性質の不明なMg-ATPaseは多数報告されている．本研究ではラット脳に存在するMg-ATPaseを分析するとともに，静脈麻酔薬の作用を検討した．ラット全脳から，Pottorfの方法に従って，形質膜（PII）とミクロソーム（PIII）分画を得た．Mg-ATPase活性の至適pHはPIIが約9.4，PIIIは約7.4であった．PIIのMg-ATPase活性はF型ATPaseの阻害剤であるNaN3によって約80%抑制され，PIIIのMg-ATPase活性はP型ATPase及びV型ATPaseの阻害剤であるNa3VO4及びBafilomycine A1によってそれぞれ約10%及び30%抑制された．ウエスタンブロッティングの結果からも、F型ATPaseはPIIに，V型ATPaseはPIIIに最も多く検出された．上記阻害剤で抑制されないMg-ATPase活性をBasal Mg-ATPaseとし，各阻害剤の存在下で各ATPaseを分別して静脈麻酔薬の影響を調べた．PropofolはV型ATPase活性、PII及びPIII Basal Mg-ATPase活性を濃度依存的に抑制したが，F型ATPase活性は80%程度活性化した．PentobarbitalはすべてのATPase活性を濃度依存的に抑制した．ThiopentalもすべてのATPase活性を抑制したが，その程度は各ATPaseによって異なった．以上の結果から、静脈麻酔薬はラット脳Mg-ATPase活性を基本的に抑制するが，Propofolはミトコンドリアに特異的な作用を及ぼすことが示唆された.
Advisors/Committee Members: 藤澤, 俊明, 鈴木, 邦明, 舩橋, 誠.
Subjects/Keywords: 静脈麻酔薬; F型ATPase; V型 ATPase; Basal Mg2+ -ATPase
Record Details
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❌
APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
宮本, . (2014). ラット脳各種ATPase活性に対する静脈麻酔薬の作用. (Doctoral Dissertation). Hokkaido University. Retrieved from http://hdl.handle.net/2115/58182
Chicago Manual of Style (16th Edition):
宮本, 健志. “ラット脳各種ATPase活性に対する静脈麻酔薬の作用.” 2014. Doctoral Dissertation, Hokkaido University. Accessed April 17, 2021.
http://hdl.handle.net/2115/58182.
MLA Handbook (7th Edition):
宮本, 健志. “ラット脳各種ATPase活性に対する静脈麻酔薬の作用.” 2014. Web. 17 Apr 2021.
Vancouver:
宮本 . ラット脳各種ATPase活性に対する静脈麻酔薬の作用. [Internet] [Doctoral dissertation]. Hokkaido University; 2014. [cited 2021 Apr 17].
Available from: http://hdl.handle.net/2115/58182.
Council of Science Editors:
宮本 . ラット脳各種ATPase活性に対する静脈麻酔薬の作用. [Doctoral Dissertation]. Hokkaido University; 2014. Available from: http://hdl.handle.net/2115/58182
Loyola University Chicago
7.
Raguimova, Olga N.
A Discrete Loop in SERCA N-Domain Plays a Role in SERCA
Headpiece Dynamics and Function.
Degree: PhD, Molecular
Biology, 2018, Loyola University Chicago
URL: https://ecommons.luc.edu/luc_diss/2842
► The sarco/endoplasmic reticulum calcium ATPase (SERCA) is the major regulator of Ca2+ levels in the cell. Deficient calcium handling in the heart has been…
(more)
▼ The sarco/endoplasmic reticulum
calcium ATPase (SERCA) is the major regulator of Ca2+ levels in the
cell. Deficient calcium handling in the heart has been linked to
heart failure, a leading cause of death in developed countries. As
of today, targeting SERCA to enhance cardiac function has not been
successful due to lack of details about SERCA structural dynamics
during Ca2+ transport. In my research, I utilized
MD simulations and variety of physical assays to determine the role
of Nβ5-β6 loop in regulation of SERCA structural dynamics during
Ca2+ transport. Previous MD simulations by our lab predicted that
the Nβ5-β6 loop regulates a SERCA structural transition from an
open to a closed conformation. Here, I showed that rationally
designed mutations of three acidic residues of the Nβ5-β6 loop
decrease SERCA headpiece closed conformation in silico and in live
cells, proving that the Nβ5-β6 loop facilitates SERCA structural
dynamics. I provided evidence that the mutated transporter is able
to hydrolyze ATP without a change in Ca2+ sensitivity, but with
significantly reduced maximal activity. I propose that the
decreased transporter activity is the direct result of the deficit
in structural dynamics regulated by the Nβ5-β6
loop. FRET measurements of the fluorescently
labeled SERCA during Ca2+ transport in vitro and ex vivo revealed
an open SERCA conformation during transient Ca2+ elevations, which
characterizes steady-state population of SERCA in the rate-limiting
step of the Ca2+ cycle. Sustained Ca2+ elevation resulted in the
accumulation of wild-type SERCA in a non-physiological high-Ca2+
affinity closed conformation. In contrast, the mutated transporter
accumulated in an open conformation due to a deficit in the
kinetics of the cytosolic headpiece closure. This
study provided insights into SERCA structural rearrangements during
the Ca2+ transport and demonstrated the importance of the Nβ5-β6
loop in SERCA structural dynamics and transporter function. These
insights could be relevant in rational design of novel therapeutics
aimed to improve cardiac function by targeting specific SERCA
conformations during rate-limiting steps of Ca2+ transport
cycle.
Subjects/Keywords: ATPase; calcium; SERCA; Molecular Biology
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APA (6th Edition):
Raguimova, O. N. (2018). A Discrete Loop in SERCA N-Domain Plays a Role in SERCA
Headpiece Dynamics and Function. (Doctoral Dissertation). Loyola University Chicago. Retrieved from https://ecommons.luc.edu/luc_diss/2842
Chicago Manual of Style (16th Edition):
Raguimova, Olga N. “A Discrete Loop in SERCA N-Domain Plays a Role in SERCA
Headpiece Dynamics and Function.” 2018. Doctoral Dissertation, Loyola University Chicago. Accessed April 17, 2021.
https://ecommons.luc.edu/luc_diss/2842.
MLA Handbook (7th Edition):
Raguimova, Olga N. “A Discrete Loop in SERCA N-Domain Plays a Role in SERCA
Headpiece Dynamics and Function.” 2018. Web. 17 Apr 2021.
Vancouver:
Raguimova ON. A Discrete Loop in SERCA N-Domain Plays a Role in SERCA
Headpiece Dynamics and Function. [Internet] [Doctoral dissertation]. Loyola University Chicago; 2018. [cited 2021 Apr 17].
Available from: https://ecommons.luc.edu/luc_diss/2842.
Council of Science Editors:
Raguimova ON. A Discrete Loop in SERCA N-Domain Plays a Role in SERCA
Headpiece Dynamics and Function. [Doctoral Dissertation]. Loyola University Chicago; 2018. Available from: https://ecommons.luc.edu/luc_diss/2842
Cornell University
8.
Chen, Cheng.
Pch2 Is A Hexameric Ring Atpase That Remodels The Meiotic Chromosome Axis Protein Hop1.
Degree: PhD, Genetics, 2014, Cornell University
URL: http://hdl.handle.net/1813/37032
► In most organisms, the accurate segregation of chromosomes during the first meiotic division requires at least one crossover between each pair of homologous chromosomes. Crossovers…
(more)
▼ In most organisms, the accurate segregation of chromosomes during the first meiotic division requires at least one crossover between each pair of homologous chromosomes. Crossovers form in meiosis from programmed double-strand breaks (DSBs) that are preferentially repaired using the homologous chromosome as a template. The PCH2 gene of budding yeast is required to establish proper meiotic chromosome structure, and to regulate meiotic DSB repair outcomes. PCH2 was also found to promote meiotic checkpoint functions, and to maintain ribosomal DNA stability during meiosis. The major focus of my thesis research has been to elucidate the molecular mechanism of Pch2 function. Pch2 contains an AAA (ATPases Associated with diverse cellular Activities) domain and is conserved in worms, fruit flies, and mammals. I performed the first detailed biochemical analysis of Pch2, and found that purified Pch2 oligomerizes into single hexameric rings in the presence of nucleotide. In addition, I showed that Pch2 directly binds to Hop1, a critical component of the synaptonemal complex that facilitates DSB repair to form crossovers. Interestingly, Hop1 binding by Pch2 induces large conformational changes in Pch2 hexamers, suggesting that Pch2 hexamers exert mechanical forces on Hop1. Importantly, I demonstrate that Pch2 subunits coordinate their ATP hydrolysis activities to displace Hop1 from large DNA substrates, providing an explanation for the altered localization of Hop1 in pch2[DELTA] mutants that was previously observed. Based on these results and other genetic and cell biological evidences I propose that Pch2 impacts multiple meiotic chromosome functions by directly regulating Hop1 localization. The second part of my thesis involves analyzing the pro-crossover Msh4-Msh5 complex, which facilitates interhomolog crossover formation by stabilizing recombination intermediates. To analyze Msh4-Msh5 function, I assayed spore viability and crossover levels for 57 msh4 and msh5 mutants and identified threshold mutants that showed wild-type spore viability but significantly decreased crossover levels. These findings suggest that a buffering mechanism exists to ensure the obligate crossover when overall crossover levels are reduced.
Advisors/Committee Members: Alani, Eric (chair), Peters, Joseph E. (committee member), Goldberg, Michael Lewis (committee member).
Subjects/Keywords: meiosis; AAA proteins; hexameric ATPase
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APA (6th Edition):
Chen, C. (2014). Pch2 Is A Hexameric Ring Atpase That Remodels The Meiotic Chromosome Axis Protein Hop1. (Doctoral Dissertation). Cornell University. Retrieved from http://hdl.handle.net/1813/37032
Chicago Manual of Style (16th Edition):
Chen, Cheng. “Pch2 Is A Hexameric Ring Atpase That Remodels The Meiotic Chromosome Axis Protein Hop1.” 2014. Doctoral Dissertation, Cornell University. Accessed April 17, 2021.
http://hdl.handle.net/1813/37032.
MLA Handbook (7th Edition):
Chen, Cheng. “Pch2 Is A Hexameric Ring Atpase That Remodels The Meiotic Chromosome Axis Protein Hop1.” 2014. Web. 17 Apr 2021.
Vancouver:
Chen C. Pch2 Is A Hexameric Ring Atpase That Remodels The Meiotic Chromosome Axis Protein Hop1. [Internet] [Doctoral dissertation]. Cornell University; 2014. [cited 2021 Apr 17].
Available from: http://hdl.handle.net/1813/37032.
Council of Science Editors:
Chen C. Pch2 Is A Hexameric Ring Atpase That Remodels The Meiotic Chromosome Axis Protein Hop1. [Doctoral Dissertation]. Cornell University; 2014. Available from: http://hdl.handle.net/1813/37032
Vanderbilt University
9.
Fischer, Rachel Anne.
Significance of Potassium Homeostasis for Neurodegeneration in Glaucoma.
Degree: PhD, Pharmacology, 2019, Vanderbilt University
URL: http://hdl.handle.net/1803/14080
► Vision loss during glaucoma results from degeneration of retinal ganglion cell (RGC) axons. Glaucoma produces deficits in axon transport in RGCs prior to structural degeneration,…
(more)
▼ Vision loss during glaucoma results from degeneration of retinal ganglion cell (RGC) axons. Glaucoma produces deficits in axon transport in RGCs prior to structural degeneration, representing a potential therapeutic window to interrupt axonopathy and prevent irreversible vision loss. Furthermore, studies indicate that axon transport deficits are accompanied by electrophysiological impairment. Electrophysiological studies in the Microbead Occlusion Model of murine glaucoma reveal that RGCs with deficits in axon transport have a reduced ability to maintain spiking frequency that arises from impaired membrane repolarization. This repolarization phenotype arises from reduced cation flux and K+ dyshomeostasis that accompanies pressure-induced decreases in Na/K-
ATPase expression and activity. In vitro studies with purified RGCs indicate that elevated pressure induces early internalization of Na/K-
ATPase that, when reversed, stabilizes cation flux and prevents K+ dyshomeostasis. Furthermore, pharmacological inhibition of the Na/K-
ATPase is sufficient to replicate pressure-induced cation influx and repolarization phase phenotypes in healthy RGCs. Our findings identify a failure to maintain electrochemical gradients and cation dyshomeostasis as an early phenotype of glaucomatous pathology in RGCs.
Advisors/Committee Members: Yaqiong Xu (committee member), Charles D. Weaver (committee member), David J. Calkins (committee member), Rebecca M. Sappington (committee member), Jerod S. Denton (Committee Chair).
Subjects/Keywords: Retina; Na/K-ATPase; Glaucoma
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APA ·
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MLA ·
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CSE |
Export
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Manager
APA (6th Edition):
Fischer, R. A. (2019). Significance of Potassium Homeostasis for Neurodegeneration in Glaucoma. (Doctoral Dissertation). Vanderbilt University. Retrieved from http://hdl.handle.net/1803/14080
Chicago Manual of Style (16th Edition):
Fischer, Rachel Anne. “Significance of Potassium Homeostasis for Neurodegeneration in Glaucoma.” 2019. Doctoral Dissertation, Vanderbilt University. Accessed April 17, 2021.
http://hdl.handle.net/1803/14080.
MLA Handbook (7th Edition):
Fischer, Rachel Anne. “Significance of Potassium Homeostasis for Neurodegeneration in Glaucoma.” 2019. Web. 17 Apr 2021.
Vancouver:
Fischer RA. Significance of Potassium Homeostasis for Neurodegeneration in Glaucoma. [Internet] [Doctoral dissertation]. Vanderbilt University; 2019. [cited 2021 Apr 17].
Available from: http://hdl.handle.net/1803/14080.
Council of Science Editors:
Fischer RA. Significance of Potassium Homeostasis for Neurodegeneration in Glaucoma. [Doctoral Dissertation]. Vanderbilt University; 2019. Available from: http://hdl.handle.net/1803/14080
University of Toronto
10.
Tuhman-Mushkin, Jana.
Structural Studies of Saccharomyces cerevisiae V1-ATPase in the Stationary Phase of Yeast Cell Culture.
Degree: 2012, University of Toronto
URL: http://hdl.handle.net/1807/32635
► Vacuolar-type ATPases (V-ATPases) are ubiquitous membrane-bound protein complexes present in the endo-membrane system of all eukaryotic cells. In eukaryotic cells, the reversible dissociation of the…
(more)
▼ Vacuolar-type ATPases (V-ATPases) are ubiquitous membrane-bound protein complexes present in the endo-membrane system of all eukaryotic cells. In eukaryotic cells, the reversible dissociation of the V1 and Vo regions is an essential mechanism for regulating V-ATPase activity. Therefore, knowledge of the structure of the dissociated V1-ATPase is necessary for understanding the regulation of V-ATPase activity. In this thesis, I showed that by introducing a 3xFLAG tag at the C terminus of different V1-ATPase subunits, highly purified V1-ATPase complex could be isolated. Electron cryomicroscopy (cryo-EM) was used for initial analysis of the intact V1-ATPase. In addition to the intact complex, partial V1-ATPase subcomplexes with different subunit compositions were isolated from yeast cells in late log phase. All of the isolated subcomplexes were found to contain the major V1-ATPase subunits A and B, but differed in the peripheral stalk subunit composition.
MAST
Advisors/Committee Members: Rubinstein, John, Biochemistry.
Subjects/Keywords: cryo-EM; V-ATPase
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APA ·
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MLA ·
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CSE |
Export
to Zotero / EndNote / Reference
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APA (6th Edition):
Tuhman-Mushkin, J. (2012). Structural Studies of Saccharomyces cerevisiae V1-ATPase in the Stationary Phase of Yeast Cell Culture. (Masters Thesis). University of Toronto. Retrieved from http://hdl.handle.net/1807/32635
Chicago Manual of Style (16th Edition):
Tuhman-Mushkin, Jana. “Structural Studies of Saccharomyces cerevisiae V1-ATPase in the Stationary Phase of Yeast Cell Culture.” 2012. Masters Thesis, University of Toronto. Accessed April 17, 2021.
http://hdl.handle.net/1807/32635.
MLA Handbook (7th Edition):
Tuhman-Mushkin, Jana. “Structural Studies of Saccharomyces cerevisiae V1-ATPase in the Stationary Phase of Yeast Cell Culture.” 2012. Web. 17 Apr 2021.
Vancouver:
Tuhman-Mushkin J. Structural Studies of Saccharomyces cerevisiae V1-ATPase in the Stationary Phase of Yeast Cell Culture. [Internet] [Masters thesis]. University of Toronto; 2012. [cited 2021 Apr 17].
Available from: http://hdl.handle.net/1807/32635.
Council of Science Editors:
Tuhman-Mushkin J. Structural Studies of Saccharomyces cerevisiae V1-ATPase in the Stationary Phase of Yeast Cell Culture. [Masters Thesis]. University of Toronto; 2012. Available from: http://hdl.handle.net/1807/32635
11.
Fraqueza, Gil.
Interação de oxometalatos de vanádio, nióbio, tungsténio e molibdénio com a Ca2+-ATPase de retículo sarcoplasmático: um alvo de ação de fármaco.
Degree: 2013, RCAAP
URL: http://www.rcaap.pt/detail.jsp?id=oai:sapientia.ualg.pt:10400.1/4829
► Tese de dout., Ciências Biológicas, especialidade em Bioquímica Inorgânica, Faculdade de Ciências e Tecnologia da Universidade do Algarve, 2013
Os oxometalatos são compostos que têm…
(more)
▼ Tese de dout., Ciências Biológicas, especialidade em Bioquímica Inorgânica, Faculdade de Ciências e Tecnologia da
Universidade do Algarve, 2013
Os oxometalatos são compostos que têm sido descritos por apresentarem propriedades
insulino-miméticas, antitumorais, antibióticas, além de serem inibidores das ATPases,
nomeadamente a Ca2+-ATPase. Contudo, os efeitos da interação dos oxometalatos com a
Ca2+-ATPase, não é um assunto completamente esclarecido.
Este estudo teve como objetivo, esclarecer o modo de atuação de oxometalatos de vanádio,
nióbio, tungsténio e molibdénio na função e estrutura da Ca2+-ATPase de RS, combinando-se
estudos cinéticos com técnicas espetroscópicas. Verificou-se que os oxometalatos estudados
inibem a atividade da Ca2+-ATPase, sendo o inibidor mais potente o decavanadato com um
IC50 de 15 µM. Foi observado que decavanadato e decaniobato inibem a Ca2+-ATPase de
modo não competitivo. Por EAA, observou-se que V10 liga-se às conformações E1, E1P, E2 e
E2P, enquanto foi confirmado que V1 liga-se preferencialmente às conformações E2 e E2P.
Por outro lado, o vanadato e o decavanadato induzem a oxidação de cisteínas na proteína. A
presença do antioxidante quercetina previne a oxidação das cisteínas, mas não a inibição da
Ca2+-ATPase por vanadato ou decavanadato. O sinal de V(IV), observado no espetro de RPE
de decavanadato na presença Ca2+-ATPase de retículo sarcoplasmático e ATP, sugere uma
redução de vanadato, devido à interação de V10 com a proteína. Estes resultados sugerem que
a maior capacidade do V10 para inibir a Ca2+-ATPase pode estar, pelo menos em parte,
relacionada com o processo de redução do vanadato associada à oxidação das cisteínas na
proteína. Finalmente estes resultados contribuem para a compreensão e aplicação de
oxometalatos e polioxometalatos como moduladores eficazes de muitos processos biológicos,
particularmente aqueles associados com homeostasia do cálcio. De facto os oxometalatos, em
especial os polioxometalatos apresentam capacidade inibitória sobre as bombas iónicas, em
especial na Ca2+-ATPase, semelhante a fármacos que têm como alvo de ação terapêutica estas
bombas iónicas. Sendo por isso de considerar em estudos, a inclusão deste tipo de
oxometalatos em fármacos com aplicação terapêutica.
Advisors/Committee Members: Aureliano, M..
Subjects/Keywords: Oxometalatos; Calcio ATPase; Decavanadate; Decaniobato
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APA (6th Edition):
Fraqueza, G. (2013). Interação de oxometalatos de vanádio, nióbio, tungsténio e molibdénio com a Ca2+-ATPase de retículo sarcoplasmático: um alvo de ação de fármaco. (Thesis). RCAAP. Retrieved from http://www.rcaap.pt/detail.jsp?id=oai:sapientia.ualg.pt:10400.1/4829
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Chicago Manual of Style (16th Edition):
Fraqueza, Gil. “Interação de oxometalatos de vanádio, nióbio, tungsténio e molibdénio com a Ca2+-ATPase de retículo sarcoplasmático: um alvo de ação de fármaco.” 2013. Thesis, RCAAP. Accessed April 17, 2021.
http://www.rcaap.pt/detail.jsp?id=oai:sapientia.ualg.pt:10400.1/4829.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Fraqueza, Gil. “Interação de oxometalatos de vanádio, nióbio, tungsténio e molibdénio com a Ca2+-ATPase de retículo sarcoplasmático: um alvo de ação de fármaco.” 2013. Web. 17 Apr 2021.
Vancouver:
Fraqueza G. Interação de oxometalatos de vanádio, nióbio, tungsténio e molibdénio com a Ca2+-ATPase de retículo sarcoplasmático: um alvo de ação de fármaco. [Internet] [Thesis]. RCAAP; 2013. [cited 2021 Apr 17].
Available from: http://www.rcaap.pt/detail.jsp?id=oai:sapientia.ualg.pt:10400.1/4829.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Fraqueza G. Interação de oxometalatos de vanádio, nióbio, tungsténio e molibdénio com a Ca2+-ATPase de retículo sarcoplasmático: um alvo de ação de fármaco. [Thesis]. RCAAP; 2013. Available from: http://www.rcaap.pt/detail.jsp?id=oai:sapientia.ualg.pt:10400.1/4829
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
University of Toronto
12.
Nano, Nardin.
Functional and Structural Characterization of human Rvb1 and Rvb2 and Screening for Inhibitors of their ATPase Activity.
Degree: PhD, 2018, University of Toronto
URL: http://hdl.handle.net/1807/102947
► Rvb1 and Rvb2 are highly conserved and essential eukaryotic AAA+ proteins linked to a wide range of cellular processes. AAA+ proteins are ATPases associated with…
(more)
▼ Rvb1 and Rvb2 are highly conserved and essential eukaryotic AAA+ proteins linked to a wide range of cellular processes. AAA+ proteins are ATPases associated with diverse cellular activities. Both proteins have been found to be part of critical cellular complexes and processes in many different species: the chromatin remodeling complexes Ino80 and SWR-C, the telomerase complex, mitotic spindle assembly, as well as phosphatidylinsitol 3-kinase-related protein kinase (PIKK) signaling. Rvb1 and Rvb2 are linked to DNA damage sensing and repair, transcriptional regulation and cell growth, therefore, implicating them in carcinogenesis. Due to wide range of Rvb1 and Rvb2 involvement, their pivotal roles and overexpression in several cancers, investigating their molecular structure and activity will aid in understanding their physiological function and explain their multifaceted role. This project sheds light on the structural and functional characteristics of human Rvb1 and Rvb2 that have been highly controversial as per several independent studies.
Rvb1 and Rvb2 were shown to form a hetero-hexameric ring complex that exhibits enhanced ATPase activity in comparison to the individual proteins, with WB motif of both subunits being essential for activity. Our investigation of human Rvbs’ oligomerization using multiple techniques has shown that an N-terminal tag induced dodecamerization of human Rvb1/Rvb2 complex and affected the oligomerization of human Rvb2 alone. A mutation in the DLLDR motif was found to cause a significant enhancement in the ATPase activity of human Rvb2.
In addition, our high-throughput screen for inhibitors of the ATPase activity of human Rvbs led to the discovery of sorafenib as a weak inhibitor of human Rvb2 and human Rvb1/2 complex. Sorafenib was characterized to be a mixed-non competitive inhibitor of human Rvb2 with an IC50 of 3.1 µM and Kd of ~ 22 µM. Our work suggests that sorafenib binds to human Rvb2 through the insertion domain (DII) as shown by SPR and ATPase assays, and that it possibly has a physiological role in the SHQ1-DKC1 disassembly.
This work advances our understanding of human Rvb1 and Rvb2 and lays out a strong foundation for targeting their ATPase activity as a strategy for the identification of novel cancer therapeutics.
2020-11-19 00:00:00
Advisors/Committee Members: Houry, Walid A., Biochemistry.
Subjects/Keywords: ATPase; Rvb1; Rvb2; 0487
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APA ·
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MLA ·
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APA (6th Edition):
Nano, N. (2018). Functional and Structural Characterization of human Rvb1 and Rvb2 and Screening for Inhibitors of their ATPase Activity. (Doctoral Dissertation). University of Toronto. Retrieved from http://hdl.handle.net/1807/102947
Chicago Manual of Style (16th Edition):
Nano, Nardin. “Functional and Structural Characterization of human Rvb1 and Rvb2 and Screening for Inhibitors of their ATPase Activity.” 2018. Doctoral Dissertation, University of Toronto. Accessed April 17, 2021.
http://hdl.handle.net/1807/102947.
MLA Handbook (7th Edition):
Nano, Nardin. “Functional and Structural Characterization of human Rvb1 and Rvb2 and Screening for Inhibitors of their ATPase Activity.” 2018. Web. 17 Apr 2021.
Vancouver:
Nano N. Functional and Structural Characterization of human Rvb1 and Rvb2 and Screening for Inhibitors of their ATPase Activity. [Internet] [Doctoral dissertation]. University of Toronto; 2018. [cited 2021 Apr 17].
Available from: http://hdl.handle.net/1807/102947.
Council of Science Editors:
Nano N. Functional and Structural Characterization of human Rvb1 and Rvb2 and Screening for Inhibitors of their ATPase Activity. [Doctoral Dissertation]. University of Toronto; 2018. Available from: http://hdl.handle.net/1807/102947
University of New South Wales
13.
Laming, Elise.
Investigating the proton-transolcating subunit of the rotary A-type ATPase.
Degree: Victor Chang Cardiac Research Institute, 2016, University of New South Wales
URL: http://handle.unsw.edu.au/1959.4/56389
;
https://unsworks.unsw.edu.au/fapi/datastream/unsworks:40642/SOURCE02?view=true
► All forms of life are dependent on proton gradients as a source of energy to power essential cellular processes, and they all store this energy…
(more)
▼ All forms of life are dependent on proton gradients as a source of energy to power essential cellular processes, and they all store this energy in the form of the nucleotide adenosine triphosphate (ATP). The rotary ATPases are multi-subunit molecular machines that utilise a rotary catalytic mechanism to couple ATP synthesis/hydrolysis to proton translocation across biological membranes. All rotary ATPases contain membrane-embedded proton-translocating subunits, via which protons enter and exit the membrane. This is the only subunit common to all rotary ATPases for which there is no high resolution structural information, and thus the molecular understanding of mechanism by which proton translocation is coupled to ATP synthesis/hydrolysis remains incomplete. To determine this structure, twelve homologues of the proton translocating subunit from A-type rotary ATPases (subunit I) were cloned into eight different expression vectors, giving 96 expression constructs. Subunit I from the bacterium Meiothermus ruber gave the highest expression. Purification of this construct was optimised and purified protein was subjected to multi-angle laser light scattering (MALLS) analysis and crystallisation trials. The interaction between subunit I and the peripheral stalks, one of the binding partners of subunit I in the intact A-
ATPase complex, was also investigated by pulldown analysis, MALLS and isothermal titration calorimetry (ITC). The X-ray crystal structure of subunit I was not able to be determined, and it was proposed that this was most likely because subunit I is most stable within the intact A
ATPase complex and in the lipid bilayer of the native organism. Nevertheless, it was found that the recombinantly expressed and purified M. ruber subunit I was able to bind to one peripheral stalk heterodimer, indicating subunit I was folded. There are, however, two peripheral stalk binding sites within subunit I, suggesting the two sites have different affinities for the peripheral stalk heterodimers. This has implications for the assembly of the intact A-
ATPase complex.
Advisors/Committee Members: Stock, Daniela, Victor Chang Cardiac Research Institute, Faculty of Medicine, UNSW, Martinac, Boris, Victor Chang Cardiac Research Institute, Faculty of Medicine, UNSW.
Subjects/Keywords: Respiration; ATPase; Proton translocating
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APA ·
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Export
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APA (6th Edition):
Laming, E. (2016). Investigating the proton-transolcating subunit of the rotary A-type ATPase. (Doctoral Dissertation). University of New South Wales. Retrieved from http://handle.unsw.edu.au/1959.4/56389 ; https://unsworks.unsw.edu.au/fapi/datastream/unsworks:40642/SOURCE02?view=true
Chicago Manual of Style (16th Edition):
Laming, Elise. “Investigating the proton-transolcating subunit of the rotary A-type ATPase.” 2016. Doctoral Dissertation, University of New South Wales. Accessed April 17, 2021.
http://handle.unsw.edu.au/1959.4/56389 ; https://unsworks.unsw.edu.au/fapi/datastream/unsworks:40642/SOURCE02?view=true.
MLA Handbook (7th Edition):
Laming, Elise. “Investigating the proton-transolcating subunit of the rotary A-type ATPase.” 2016. Web. 17 Apr 2021.
Vancouver:
Laming E. Investigating the proton-transolcating subunit of the rotary A-type ATPase. [Internet] [Doctoral dissertation]. University of New South Wales; 2016. [cited 2021 Apr 17].
Available from: http://handle.unsw.edu.au/1959.4/56389 ; https://unsworks.unsw.edu.au/fapi/datastream/unsworks:40642/SOURCE02?view=true.
Council of Science Editors:
Laming E. Investigating the proton-transolcating subunit of the rotary A-type ATPase. [Doctoral Dissertation]. University of New South Wales; 2016. Available from: http://handle.unsw.edu.au/1959.4/56389 ; https://unsworks.unsw.edu.au/fapi/datastream/unsworks:40642/SOURCE02?view=true
Vanderbilt University
14.
Takar, Mehmet.
Regulation of membrane asymmetry by Golgi P4-ATPases and their interactors.
Degree: PhD, Biological Sciences, 2018, Vanderbilt University
URL: http://hdl.handle.net/1803/14437
► Neo1 was discovered in a multi-copy suppressor screen in Saccharomyces cerevisiae to search for factors conferring resistance against an aminoglycoside antibiotic, neomycin (1). Aminoglycoside antibiotics,…
(more)
▼ Neo1 was discovered in a multi-copy suppressor screen in Saccharomyces cerevisiae to search for factors conferring resistance against an aminoglycoside antibiotic, neomycin (1). Aminoglycoside antibiotics, used to treat bacterial infections by interfering with proofreading during protein synthesis, cause hearing loss in genetically susceptible individuals. It remains unclear why Neo1 overexpression confers resistance to neomycin. Among the yeast P4-ATPases, Neo1 is the only essential P4-
ATPase and it has critical roles in COPI-mediated retrograde transport as well as Golgi glycosylation (2,3). In addition, neo1ts mutants display hyperacidification of Golgi membranes and this defect is tightly associated with Neo1’s role in Golgi glycosylation (4). My thesis research studies revealed that Neo1 regulates phosphatidylethanolamine (PE) and phosphatidylserine (PS) asymmetry of the plasma membrane, supporting its proposed function as a phospholipid flippase (5). I provided evidence for the regulation of plasma membrane asymmetry directly by Neo1 rather than indirectly by influencing the activity of Drs2/Dnf P4-ATPases (5). Neo1 also regulates the vacuole fusion machinery and our data suggest this occurs through the enrichment of PE on the cytosolic leaflet of the vacuole membrane (6). The Neo1 homolog Tat-5 in C. elegans was also associated with PE transport activity during embryonic development (7). Together these studies suggest PE is the primary transport substrate for the Neo1/Tat-5/ATP9 orthologs. Unlike Drs2/Dnf P4-ATPases, Neo1 and its human orthologues (Atp9a and Atp9b) do not require a Cdc50-related beta-subunit for either its ER exit nor for its function in vivo (8-11). Evidence others and we have accumulated strongly support the hypothesized role of Neo1 as a phospholipid transporter, but it is still unclear how Neo1 activity is regulated in the Golgi/endosomal membranes.
In the second part of my thesis, we discovered a novel factor antagonizing the Golgi flippase activity called Antagonizes Neo1 Yeast phospholipid flippase (ANY1) in a collaborative study with Charles Boone’s research group. We found that any1∆ completely suppresses the defects that occur from loss of Neo1 and its interaction partners Dop1 and Mon2. Our collaborative study showed that overexpression of Any1 redistributes PS pools from the plasma membrane to internal organelles. On the contrary, overexpression of Any1 only leads to the partial loss of PE asymmetry, but not PS asymmetry. I further identified an essential requirement for a PS flippase activity in the Golgi membranes in the absence of Any1. Serendipitously, I determined that a mutation in transmembrane segment 2 of Neo1 (Neo1[Y222S]) can suppress drs2∆ growth and membrane asymmetry defects. More interestingly, I revealed that the gain of function phenotype is dependent on Any1. Molecular interaction studies performed by immunoprecipitation and proteomics also determined that Neo1 and Any1 physically interact with each other in the Golgi membranes, but it is not clear whether the…
Advisors/Committee Members: Katherine L. Friedman (committee member), Kathleen L. Gould (committee member), John D. York (committee member), Todd R. Graham (committee member), Brandt Eichman (Committee Chair).
Subjects/Keywords: membrane asymmetry; P4-ATPase; ATPase; transporter; phosphatidylserine; phosphatidylethanolamine; membrane lipid; scramblase
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APA (6th Edition):
Takar, M. (2018). Regulation of membrane asymmetry by Golgi P4-ATPases and their interactors. (Doctoral Dissertation). Vanderbilt University. Retrieved from http://hdl.handle.net/1803/14437
Chicago Manual of Style (16th Edition):
Takar, Mehmet. “Regulation of membrane asymmetry by Golgi P4-ATPases and their interactors.” 2018. Doctoral Dissertation, Vanderbilt University. Accessed April 17, 2021.
http://hdl.handle.net/1803/14437.
MLA Handbook (7th Edition):
Takar, Mehmet. “Regulation of membrane asymmetry by Golgi P4-ATPases and their interactors.” 2018. Web. 17 Apr 2021.
Vancouver:
Takar M. Regulation of membrane asymmetry by Golgi P4-ATPases and their interactors. [Internet] [Doctoral dissertation]. Vanderbilt University; 2018. [cited 2021 Apr 17].
Available from: http://hdl.handle.net/1803/14437.
Council of Science Editors:
Takar M. Regulation of membrane asymmetry by Golgi P4-ATPases and their interactors. [Doctoral Dissertation]. Vanderbilt University; 2018. Available from: http://hdl.handle.net/1803/14437
15.
D'Silva, Natalie.
THE ROLE OF ION-MOTIVE ATPASES IN THE INSECT GUT.
Degree: PhD, 2018, McMaster University
URL: http://hdl.handle.net/11375/24079
► The present set of studies examines the roles of two ion-motive enzymes, vacuolar-type H+-ATPase (VA) and Na+/K+ ATPase (NKA), in energizing transepithelial ion transport across…
(more)
▼ The present set of studies examines the roles of two ion-motive enzymes, vacuolar-type H+-ATPase (VA) and Na+/K+ ATPase (NKA), in energizing transepithelial ion transport across the larval caecum and midgut epithelia of Drosophila melanogaster and Aedes aegypti. Even though both VA and NKA are expressed in insect epithelia, VA was considered the more important enzyme until the early 2000 because the ion transport was unaffected by the NKA inhibitor ouabain in many insect epithelia, a phenomenon termed the ‘ouabain paradox’. This paradox was resolved by the discovery of an organic anion transporter (OATP) that is colocalized with NKA and prevents the actions of ouabain on NKA. Since the resolution of the ouabain paradox, this is the first set of studies that investigates the role of NKA in energizing ion transport across the caeca and midgut of insects. First, I show that both VA and NKA are expressed in the caecum and the midgut. Moreover, the ATPase enzyme activities of VA and NKA are quantitatively similar within each region of the gut that was studied, suggesting that both ATPases may be important for establishing favourable electrochemical gradients for transport of ions across the gut. I used ATPase inhibitors to demonstrate that cation transport is dependent on the actions of both VA and NKA. Furthermore, this is the first set of studies that provides an insight into the ion transport mechanisms of the gastric caecum, an organ that is understudied in insects. In Aedes aegypti, I show that 5-hydroxytryptamine regulates the VA-rich cells of the gastric caecum, and therefore the rates of ion transport of these cells. Additionally, I also show that rearing salinity conditions for Aedes aegypti larvae alters the expression patterns of VA and NKA in the gastric caecum. In freshwater, increased activity of VA and NKA energizes transport of ions into the lumen of the caecum that likely maintains fluid volumes and ionic composition at levels appropriate for digestion and absorption. Overall, these studies provide novel information for caeca and midgut-specific actions of VA and NKA in insects, and present a number of new avenues for future research.
Thesis
Doctor of Philosophy (PhD)
This thesis focuses on investigating the roles of two enzymes, vacuolar-type H+-ATPase (VA) and Na+/K+ ATPase (NKA), which utilize energy to transport electrically charged atoms (ions) across the cells of the insect gut. Although VA was considered the more important of the two enzymes until the early 2000s, I have demonstrated that NKA also plays a role in maintaining insect gut function in fruit flies and mosquito larvae. Furthermore, the activities of both enzymes are dependent on the salinity of the medium in which mosquito larvae are reared, suggesting that they play a role in maintaining the ionic composition of the gut fluids in freshwater larvae. Additionally, I have also demonstrated that a neurochemical, serotonin, can modulate the activity of gut cells in mosquito larvae. Overall, this thesis provides novel information on…
Advisors/Committee Members: O'Donnell, Michael, Biology.
Subjects/Keywords: Insect; Physiology; V-ATPase; Na+/K+ ATPase; ion-transport
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APA ·
Chicago ·
MLA ·
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Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
D'Silva, N. (2018). THE ROLE OF ION-MOTIVE ATPASES IN THE INSECT GUT. (Doctoral Dissertation). McMaster University. Retrieved from http://hdl.handle.net/11375/24079
Chicago Manual of Style (16th Edition):
D'Silva, Natalie. “THE ROLE OF ION-MOTIVE ATPASES IN THE INSECT GUT.” 2018. Doctoral Dissertation, McMaster University. Accessed April 17, 2021.
http://hdl.handle.net/11375/24079.
MLA Handbook (7th Edition):
D'Silva, Natalie. “THE ROLE OF ION-MOTIVE ATPASES IN THE INSECT GUT.” 2018. Web. 17 Apr 2021.
Vancouver:
D'Silva N. THE ROLE OF ION-MOTIVE ATPASES IN THE INSECT GUT. [Internet] [Doctoral dissertation]. McMaster University; 2018. [cited 2021 Apr 17].
Available from: http://hdl.handle.net/11375/24079.
Council of Science Editors:
D'Silva N. THE ROLE OF ION-MOTIVE ATPASES IN THE INSECT GUT. [Doctoral Dissertation]. McMaster University; 2018. Available from: http://hdl.handle.net/11375/24079
16.
Grigoletto, Aude.
Rôle de la reptine dans le carcinome hépatocellulaire : Role of Reptin in hepatocellular carcinoma.
Degree: Docteur es, Sciences, technologie, santé. Biologie cellulaire et physiopathologie, 2012, Université de Bordeaux Segalen
URL: http://www.theses.fr/2012BOR21946
► Le carcinome hépatocellulaire (CHC) est le principal cancer primitif du foie et est associé à un très mauvais pronostic. Notre équipe a mis en évidence…
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▼ Le carcinome hépatocellulaire (CHC) est le principal cancer primitif du foie et est associé à un très mauvais pronostic. Notre équipe a mis en évidence que la Reptine et la Pontine, des AAA+ ATPases homologues, sont surexprimées dans le CHC par rapport au foie non tumoral. Au cours de ce travail de thèse, j’ai contribué à démontrer que l’extinction de la Reptine par l’induction de shRNA suffit à arrêter la croissance de tumeurs déjà établies, et même à induire leur régression dans des xénogreffes chez la souris. Ces résultats encourageants suggèrent que la Reptine pourrait être une cible thérapeutique dans le CHC. L’utilisation de siRNA en thérapeutique n’étant pas envisageable actuellement, il parait plus pertinent de tenter de cibler la Reptine via son activité ATPase. Le principal objectif de ma thèse était donc de déterminer l’implication de l’activité ATPase de la Reptine pour ses propriétés oncogéniques dans le CHC. Nos résultats ont montré que des mutants inactifs de la Reptine (D299N et E300G) ont un effet dominant négatif et ne sont pas capables de complémenter l’absence de la Reptine endogène, ce qui conduit à une diminution significative de la croissance des cellules HuH7 et Hep3B, et à une induction de l’apoptose. Ceci indique que l’activité ATPase de la Reptine est nécessaire pour la croissance et la survie des cellules de CHC. Enfin, grâce à une étude transcriptomique, nous avons identifié de nouveaux gènes dont l’expression est régulée par la Reptine et/ou la Pontine. Parmi ces gènes, certains pourraient être impliqués dans les fonctions oncogéniques de la Reptine et/ou de la Pontine dans le CHC. Finalement, ce travail a permis de mettre en évidence l’implication de l’activité ATPase de la Reptine, et d’apporter des éléments permettant de mieux comprendre le mécanisme d’action de la Reptine dans le CHC.
Hepatocellular carcinoma (HCC) is the main primary cancer of the liver and is often associated with poor prognosis. Our team has demonstrated that Reptin and Pontin, two AA+ ATPases, are overexpressed in HCC compared to normal liver. Moreover this overexpression is also associated with poor prognosis. In the course of my PhD, I demonstrated that shRNA-mediated silencing of Reptin is sufficient to inhibit tumor growth and even can promote their regression in xenografted mice. These encouraging results suggest that Reptin might represent a novel therapeutic target in HCC. As the use of siRNA as therapeutic tools is still debated, the targeting of Reptin enzymatic activity might represent a more relevant approach to impair its functions. To this end I first proposed to determine the involvement of Reptin ATPase activity in HCC oncogenesis. My results show that ATPase inactive Reptin mutants (D299N and E300G) play dominant negative roles toward Reptin functions and are unable to complement for the depletion of endogenous Reptin, thereby leading to a significant decrease of cell growth and to a significant increase of apoptosis in HuH7 and Hep3B cells. These results show that Reptin’s ATPase activity is…
Advisors/Committee Members: Rosenbaum, Jean (thesis director).
Subjects/Keywords: Carcinome hépatocellulaire; Reptine; Pontine; ATPase; Hepatocellular carcinoma; Reptin; Pontin; ATPase
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APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
Grigoletto, A. (2012). Rôle de la reptine dans le carcinome hépatocellulaire : Role of Reptin in hepatocellular carcinoma. (Doctoral Dissertation). Université de Bordeaux Segalen. Retrieved from http://www.theses.fr/2012BOR21946
Chicago Manual of Style (16th Edition):
Grigoletto, Aude. “Rôle de la reptine dans le carcinome hépatocellulaire : Role of Reptin in hepatocellular carcinoma.” 2012. Doctoral Dissertation, Université de Bordeaux Segalen. Accessed April 17, 2021.
http://www.theses.fr/2012BOR21946.
MLA Handbook (7th Edition):
Grigoletto, Aude. “Rôle de la reptine dans le carcinome hépatocellulaire : Role of Reptin in hepatocellular carcinoma.” 2012. Web. 17 Apr 2021.
Vancouver:
Grigoletto A. Rôle de la reptine dans le carcinome hépatocellulaire : Role of Reptin in hepatocellular carcinoma. [Internet] [Doctoral dissertation]. Université de Bordeaux Segalen; 2012. [cited 2021 Apr 17].
Available from: http://www.theses.fr/2012BOR21946.
Council of Science Editors:
Grigoletto A. Rôle de la reptine dans le carcinome hépatocellulaire : Role of Reptin in hepatocellular carcinoma. [Doctoral Dissertation]. Université de Bordeaux Segalen; 2012. Available from: http://www.theses.fr/2012BOR21946
University of Kansas
17.
He, Lucy.
Bioenergetic Regulation of Neuronal Vacuolar-type H+- ATPase.
Degree: MS, Pharmacology & Toxicology, 2019, University of Kansas
URL: http://hdl.handle.net/1808/30212
► Neuronal vacuolar-type H+-ATPase (V-ATPase) is an ATP-dependent proton pump that functions to acidify intracellular organelles such as lysosomes and synaptic vesicles, creating a proton gradient…
(more)
▼ Neuronal vacuolar-type H+-
ATPase (V-
ATPase) is an ATP-dependent proton pump that functions to acidify intracellular organelles such as lysosomes and synaptic vesicles, creating a proton gradient by which neurotransmitters can enter the vesicle through proton-coupled neurotransmitter transporters, a crucial step in neurotransmission (Moriyama, Maeda, & Futai, 1992). It is composed of two reversible domains, the integral V0 that allows proton translocation and catalytic peripheral V1 that is responsible for ATP hydrolysis. The V-
ATPase regulates its activity through a process called reversible disassembly. When the V0 and V1 domains assemble, V-
ATPase is activated and allows the influx of protons. When the domains disassemble, V-
ATPase is inactivated, and proton transport does not occur (Beltran & Nelson, 1992). V-
ATPase assembly was previously demonstrated to be regulated by glucose in yeast (Kane, 1995) and some mammalian cells (Toei, Saum, & Forgac, 2010), but whether and how glucose regulates neuronal V-
ATPase is unclear. This study investigates the effect of bioenergetic substrates on neuronal V-
ATPase assembly. Neuro2a (N2a) cells were differentiated for 96 hours, glucose-deprived overnight, and then treated with substrates such as glucose, beta-hydroxybutyrate, sodium pyruvate, creatine phosphate, and creatine monohydrate for 20 minutes prior to cell lysate preparation. To study V-
ATPase assembly, assembled V-
ATPase were captured through co-immunoprecipitation. Equal amounts of cell lysate were incubated with V1 antibody-coupled resin. Immunoblotting was then performed on the eluate to detect the V1 and V0 domains. The density of the bands was quantified and the ratio of V0 and V1 domains was used to determine V0/ V1 assembly. The results demonstrate changes in glucose availability (deprivation/stimulation) did not impact V-
ATPase assembly in differentiated N2a cells. Furthermore, the results show other bioenergetic substrates (pyruvate, beta-hydroxybutyrate, and creatine molecules) did not induce changes in neuronal V-
ATPase assembly. Our results did not support the well-documented role of glucose regulation of V-
ATPase assembly demonstrated in previous studies conducted in yeast and mammalian cells. Several pitfalls of this study may have contributed to the negative results. The glucose concentration for stimulation and incubation time for glucose-deprivation may not have been optimal for differentiated N2a cells; further investigation will be needed to optimize these conditions. The glucose concentration used for stimulation after overnight glucose-deprivation did not exceed the physiological glucose concentration; higher concentrations of glucose should be tested. Previous studies had various incubation times for glucose-deprivation that could be experimentally optimized. Furthermore, the experiments could be replicated in other neuronal cells or in primary neurons to validate the results.
Advisors/Committee Members: Zhao, Liqin (advisor), Rosa-Molinar, Eduardo (cmtemember), Moskovitz, Jackob (cmtemember).
Subjects/Keywords: Pharmacology; Bioenergetic Substrates; glucose; N2a; neuronal; V-ATPase; V-ATPase assembly
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APA ·
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MLA ·
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APA (6th Edition):
He, L. (2019). Bioenergetic Regulation of Neuronal Vacuolar-type H+- ATPase. (Masters Thesis). University of Kansas. Retrieved from http://hdl.handle.net/1808/30212
Chicago Manual of Style (16th Edition):
He, Lucy. “Bioenergetic Regulation of Neuronal Vacuolar-type H+- ATPase.” 2019. Masters Thesis, University of Kansas. Accessed April 17, 2021.
http://hdl.handle.net/1808/30212.
MLA Handbook (7th Edition):
He, Lucy. “Bioenergetic Regulation of Neuronal Vacuolar-type H+- ATPase.” 2019. Web. 17 Apr 2021.
Vancouver:
He L. Bioenergetic Regulation of Neuronal Vacuolar-type H+- ATPase. [Internet] [Masters thesis]. University of Kansas; 2019. [cited 2021 Apr 17].
Available from: http://hdl.handle.net/1808/30212.
Council of Science Editors:
He L. Bioenergetic Regulation of Neuronal Vacuolar-type H+- ATPase. [Masters Thesis]. University of Kansas; 2019. Available from: http://hdl.handle.net/1808/30212
University of California – Berkeley
18.
Gross, Sean.
The effects of oxidative modifications on cardiac and skeletal muscle.
Degree: Integrative Biology, 2011, University of California – Berkeley
URL: http://www.escholarship.org/uc/item/6k89d44x
► Biological systems are frequently exposed to reactive oxygen species (ROS), which can be damaging to cellular function. Recent research suggests that ROS can also be…
(more)
▼ Biological systems are frequently exposed to reactive oxygen species (ROS), which can be damaging to cellular function. Recent research suggests that ROS can also be used as signaling agents, modifying protein functions post-translationally, by reversibly modifying cysteines. The contractile functions of both skeletal and cardiac muscle are responsive to reactive oxygen species, but the mechanisms are unknown. We therefore developed methods to quantify, which cysteines are modified by exposure to oxidative reagents, and used these methods as a gateway for testing different mechanisms for how ROS affect function. From these techniques, we found that nearly every myofilament protein in both cardiac and skeletal muscle has one or more cysteines that can be modified by ROS in the environment of the myofilament lattice. Interestingly, we found that both the cysteines modified and the effects from ROS exposure depended on the contractile state of the muscle. Exposure in a relaxing buffer (ATP present) modified a large number of myosin thiols, and decreased the maximum ATPase and calcium sensitivity when compared to untreated myofibrils. In contrast, ROS exposure in a rigor buffer (no ATP) decreased the number of modified myosin thiols and increased the modification of actin when compared to treatment in relaxing buffer. Exposure in rigor buffer did not change the maximum myofibril ATPase, but did increase the basal ATPase and calcium sensitivity. Complementary studies in fast skeletal muscle tested the ability of different cysteine reagents to change calcium sensitivity. We found that different reagents affected calcium sensitivity in opposite directions, and found evidence that the difference in direction was due to differences in the type of modification, rather than a difference between cysteine sites. We narrowed down the proteins responsible for these effects to actin, essential light chain, tropomyosin and the myosin heavy chain. Further eliminating proteins with cysteines not unique to fast muscle, we found two cysteines in the myosin heavy chain, both in the neck region, that are the most likely to be responsible for the changes in calcium sensitivity. In summary these experiments shed significant light on a variety of mechanisms by which ROS can modify the contractile function of cardiac and skeletal muscle. These results may be important for understanding and eventually treatment of diseases that have been found to produce oxidative modifications and decreased function of muscle, including heart failure, muscular dystrophy, arthritis and cancer.
Subjects/Keywords: Physiology; ATPase; calcium sensitivity; cysteine; Muscle; ROS
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Manager
APA (6th Edition):
Gross, S. (2011). The effects of oxidative modifications on cardiac and skeletal muscle. (Thesis). University of California – Berkeley. Retrieved from http://www.escholarship.org/uc/item/6k89d44x
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Chicago Manual of Style (16th Edition):
Gross, Sean. “The effects of oxidative modifications on cardiac and skeletal muscle.” 2011. Thesis, University of California – Berkeley. Accessed April 17, 2021.
http://www.escholarship.org/uc/item/6k89d44x.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Gross, Sean. “The effects of oxidative modifications on cardiac and skeletal muscle.” 2011. Web. 17 Apr 2021.
Vancouver:
Gross S. The effects of oxidative modifications on cardiac and skeletal muscle. [Internet] [Thesis]. University of California – Berkeley; 2011. [cited 2021 Apr 17].
Available from: http://www.escholarship.org/uc/item/6k89d44x.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Gross S. The effects of oxidative modifications on cardiac and skeletal muscle. [Thesis]. University of California – Berkeley; 2011. Available from: http://www.escholarship.org/uc/item/6k89d44x
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
University of Manchester
19.
Wang, Yanwen.
Studies of Ca2+ handling and electrophysiological
properties in murine heartswith genetic modification of plasma
membrane Ca2+ ATPase 1.
Degree: 2013, University of Manchester
URL: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:184640
► In heart, Ca2+ plays an important role in maintenance of normal cardiac functions. Regulation of Ca2+ is mainly through L-type Ca2+ channel (LTCC), Ryanodine receptor…
(more)
▼ In heart, Ca2+ plays an important role in
maintenance of normal cardiac functions. Regulation of Ca2+ is
mainly through L-type Ca2+ channel (LTCC), Ryanodine receptor (RyR)
and sarcoplasmic reticulum calcium ATPase pump (SERCA) on
sarcoplasmic reticulum (SR), Na+-Ca2+ exchanger (NCX), plasma
membrane Ca2+ ATPase (PMCA). It has been well-accepted that PMCA
plays a minor contribution to elevation of Ca2+ compared to SERCA
and NCX and in regulation of cytosolic Ca2+ homeostasis. There are
four isoforms of PMCA, PMCA1-4, and PMCA1 is a house-keeping gene,
and abundantly distributed in heart. However, the role of PMCA1 in
the murine heart has not been fully explored. With a cardiac
specific knockout mouse model, the electrophysiological
characteristics of PMCA1 in murine hearts, particularly in atria
under normal physiological and stress conditions ([Ca2+]o overload
and pacing conditions) are investigated. Firstly the complete
deletion of PMCA1 in the atria in PMCA1cko mice was confirmed by
Western blotting and immunostaining, also the membrane localisation
of PMCA1 in the atria in PMCA1loxP/loxP mice was demonstrated. Then
the phenotypes of ex vivo whole hearts between PMCA1loxP/loxP and
PMCA1cko mice under physiological conditions and [Ca2+]o overload
condition and with different frequencies by programmed electrical
stimulation (PES) were explored. Further more, the Ca2+ handling
process in single atrial myocytes between the PMCA1 deletion mice
and control mice under normal physiological conditions and [Ca2+]o
overload condition and stimulation with different frequencies was
investigated. Finally the Ca2+ handling process in single
ventricular myocytes between the PMCA1 deletion mice and control
mice under normal physiological condition was investigated. At the
whole heart level, the PMCA1cko hearts became more susceptible to
arrhythmias with PES under physiological conditions compared with
the PMCA1loxP/loxP hearts, and such arrhythmic events occurred more
often and had longer pacing durations under Ca2+ overload
conditions and higher frequency of pacing. At the single cellular
level, the NCX current decay was significantly prolonged in
PMCA1cko atrial myocytes under physiological conditions. This was
further increased under Ca2+ overload conditions. With
frequency-dependent stimulation, the PMCA1cko atrial myocytes
showed few EAD- or DAD-type APs under physiological conditions in
contrast to PMCA1loxP/loxP atrial myocytes that showed no
arrhythmic events. The occurrence increased significantly under
Ca2+ overload condition and/or at higher frequency of stimulation.
Similar findings were observed in isolated ventricular myocytes. To
conclude, the role of PMCA1 in maintaining Ca2+ homeostasis and
electrical function in atrial myocytes under physiological
conditions is minor. ii) PMCA1 has a critical role in maintaining
Ca2+ homeostasis and electrical function in the atrium under stress
conditions. This is particularly important during fast efflux of
Ca2+ which is required under stress conditions.
Cardiac disease is…
Advisors/Committee Members: WANG, TAO T, Wang, Tao, Lei, Ming.
Subjects/Keywords: plasma membrane Ca2+-ATPase; cardiac myocyte; electrophysiology
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APA ·
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MLA ·
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to Zotero / EndNote / Reference
Manager
APA (6th Edition):
Wang, Y. (2013). Studies of Ca2+ handling and electrophysiological
properties in murine heartswith genetic modification of plasma
membrane Ca2+ ATPase 1. (Doctoral Dissertation). University of Manchester. Retrieved from http://www.manchester.ac.uk/escholar/uk-ac-man-scw:184640
Chicago Manual of Style (16th Edition):
Wang, Yanwen. “Studies of Ca2+ handling and electrophysiological
properties in murine heartswith genetic modification of plasma
membrane Ca2+ ATPase 1.” 2013. Doctoral Dissertation, University of Manchester. Accessed April 17, 2021.
http://www.manchester.ac.uk/escholar/uk-ac-man-scw:184640.
MLA Handbook (7th Edition):
Wang, Yanwen. “Studies of Ca2+ handling and electrophysiological
properties in murine heartswith genetic modification of plasma
membrane Ca2+ ATPase 1.” 2013. Web. 17 Apr 2021.
Vancouver:
Wang Y. Studies of Ca2+ handling and electrophysiological
properties in murine heartswith genetic modification of plasma
membrane Ca2+ ATPase 1. [Internet] [Doctoral dissertation]. University of Manchester; 2013. [cited 2021 Apr 17].
Available from: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:184640.
Council of Science Editors:
Wang Y. Studies of Ca2+ handling and electrophysiological
properties in murine heartswith genetic modification of plasma
membrane Ca2+ ATPase 1. [Doctoral Dissertation]. University of Manchester; 2013. Available from: http://www.manchester.ac.uk/escholar/uk-ac-man-scw:184640
20.
Gabriel Costa Nunes da Cruz.
Caracterização parcial de uma Ca2+ -ATPase de larva de Pachymerus nucleorum (Coleoptera: Chrysomelidae: Bruchinae).
Degree: 2006, Federal University of Uberlândia
URL: http://www.bdtd.ufu.br//tde_busca/arquivo.php?codArquivo=504
;
http://www.bdtd.ufu.br//tde_busca/arquivo.php?codArquivo=505
► A high Ca2+-ATPase activity fraction was obtained from Pachymerus nucleorum larvae. The larvae were dissected in order to take off the digestive system. The remaining…
(more)
▼ A high Ca2+-ATPase activity fraction was obtained from Pachymerus nucleorum larvae. The larvae were dissected in order to take off the digestive system. The remaining material was homogeneized in extraction buffer and centrifuged at 15000xg for 30 minutes. The resulting precipitate, fraction P1, was homogeneized in imidazole buffer containing Triton X-100 0.2%, incubated for 20 minutes at room temperature and centrifuged under the aforementioned conditions. The precipitate fraction P2 was homogeneized in imidazole buffer containing 50 mM pyrophosphate. After incubation for 20 minutes at room temperature, a centrifugation at 40000xg for 30 minutes was done. The precipitate fraction P3 was homogeneized in imidazole buffer and presented high Ca2+- ATPase activity that was characterized. The fraction P3 did not present significant ATPase activity in presence of some cations: magnesium, cobalt, cupper, zinc, barium, lithium or iron. The high Ca2+-ATPase activity was inhibited in presence of 4 mM cobalt, cupper, zinc, barium or iron. 0.25 mM magnesium caused 50% inhibition of this activity. In presence of 1 mM ATP the P3 ATPase activity gets 50% with 0.5 mM CaCl2, reaching a plateau with 2 mM. In presence of 1 mM 31 CaCl2 the ATPase activity gets 50% with 0.7 mM ATP and the pick with 2 mM. However we observed that there is a slight decline of this activity with ATP concentrations higher than 2 mM. P3 used neither pyrophosphate nor mono- or diphosphate nucleotides as substrate, and the Ca2+-GTPase activity is only 20% that of the Ca2+-ATPase. This fraction did not undergo significant inhibition by vanadate, in concentrations smaller than 200 M, azide, Triton X-100 or aluminum fluoride. The K+/EDTA-ATPase activity was 50% of the Ca2+-ATPase and did not occur imunoreactivity with anti-myosin II or V antibodies. In this work we obtained a Ca2+-ATPase activity enriched fraction from Pachymerus nucleorum larvae lacking digestive system and partially characterized this activity.
RESUMO GERAL - O babaçu (Orbignya sp.) é uma palmeira que ocupa vastas áreas na região meio-norte do Brasil e desempenha um papel importante na subsistência de milhares de habitantes desta região. Pachymerus nucleorum é um besouro da subfamília Bruchinae, cujos representantes são conhecidos como brocas de sementes, que causa muitos danos nas castanhas do babaçu. As larvas deste animal penetram o coco de babaçu logo após sua eclosão, se desenvolvem alimentando-se das castanhas e somente emergem como adultos. Neste trabalho, estudamos a atividade enzimática de hidrólise de ATP por uma fração protéica parcialmente purificada a partir das larvas de P. nucleorum sem o aparelho digestivo. As larvas foram extraídas dos cocos, lavadas e dissecadas para a retirada do aparelho digestivo. Seguimos o processo de purificação a partir do restante do corpo e obtivemos a fração precipitada (P3) com alta atividade ATPásica em presença de cálcio, que foi caracterizada. A dosagem da atividade ATPásica foi realizada incubando-se amostras das frações em meio de reação…
Advisors/Committee Members: Milton Vieira Coelho, Hérica de Lima Santos, Pietro Ciancaglini.
Subjects/Keywords: GENETICA; Bruchinae; Ca2+ -ATPase; Pachymerus nucleorum
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APA (6th Edition):
Cruz, G. C. N. d. (2006). Caracterização parcial de uma Ca2+ -ATPase de larva de Pachymerus nucleorum (Coleoptera: Chrysomelidae: Bruchinae). (Thesis). Federal University of Uberlândia. Retrieved from http://www.bdtd.ufu.br//tde_busca/arquivo.php?codArquivo=504 ; http://www.bdtd.ufu.br//tde_busca/arquivo.php?codArquivo=505
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Chicago Manual of Style (16th Edition):
Cruz, Gabriel Costa Nunes da. “Caracterização parcial de uma Ca2+ -ATPase de larva de Pachymerus nucleorum (Coleoptera: Chrysomelidae: Bruchinae).” 2006. Thesis, Federal University of Uberlândia. Accessed April 17, 2021.
http://www.bdtd.ufu.br//tde_busca/arquivo.php?codArquivo=504 ; http://www.bdtd.ufu.br//tde_busca/arquivo.php?codArquivo=505.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Cruz, Gabriel Costa Nunes da. “Caracterização parcial de uma Ca2+ -ATPase de larva de Pachymerus nucleorum (Coleoptera: Chrysomelidae: Bruchinae).” 2006. Web. 17 Apr 2021.
Vancouver:
Cruz GCNd. Caracterização parcial de uma Ca2+ -ATPase de larva de Pachymerus nucleorum (Coleoptera: Chrysomelidae: Bruchinae). [Internet] [Thesis]. Federal University of Uberlândia; 2006. [cited 2021 Apr 17].
Available from: http://www.bdtd.ufu.br//tde_busca/arquivo.php?codArquivo=504 ; http://www.bdtd.ufu.br//tde_busca/arquivo.php?codArquivo=505.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Cruz GCNd. Caracterização parcial de uma Ca2+ -ATPase de larva de Pachymerus nucleorum (Coleoptera: Chrysomelidae: Bruchinae). [Thesis]. Federal University of Uberlândia; 2006. Available from: http://www.bdtd.ufu.br//tde_busca/arquivo.php?codArquivo=504 ; http://www.bdtd.ufu.br//tde_busca/arquivo.php?codArquivo=505
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
University of Toronto
21.
Johnson, Lisa E.
R740S Mutation of Tcirg1 Mutation Affects Enamel Development in Osteopetrotic Mice.
Degree: 2016, University of Toronto
URL: http://hdl.handle.net/1807/74943
► Abstract Osteopetrosis is characterized by sclerotic bone due to impaired osteoclasts. Many cases are associated with defects in the TCIRG1 gene encoding the a3 subunit…
(more)
▼ Abstract
Osteopetrosis is characterized by sclerotic bone due to impaired osteoclasts.
Many cases are associated with defects in the TCIRG1 gene encoding the a3
subunit of vacuolar-ATPase (V-ATPase). These patients have defects in bone,
tooth eruption and enamel. To elucidate the role of V-ATPases in amelogenesis,
we investigated the cellular distribution of a3, expression of enamel proteins,
and quality of the enamel in a mouse with a mutation (R740S) in a3. In the
homozygote (R740S/R740S), micro CT and SEM demonstrated a decrease in
mineralization and thickness of the enamel. Using immunohistochemistry, the
expression of the secretory stage protein, amelogenin remained in the
R740S/R740S ameloblasts at day 9, concurrent with lower levels of the
maturation stage proteins, amelotin and ODAM, suggesting a developmental
delay. Low level expression of a3 was observed in the ameloblasts during the
secretory stage. Our results suggest that a3-V-ATPases may play a role in
amelogenesis, possibly by affecting protein secretion.
M.Sc.
Advisors/Committee Members: Voronov, Irina, Bradley, Grace, Dentistry.
Subjects/Keywords: Amelogenesis; Enamel; Osteopetrosis; Tcirg1; vacuolar ATPase; 0567
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APA (6th Edition):
Johnson, L. E. (2016). R740S Mutation of Tcirg1 Mutation Affects Enamel Development in Osteopetrotic Mice. (Masters Thesis). University of Toronto. Retrieved from http://hdl.handle.net/1807/74943
Chicago Manual of Style (16th Edition):
Johnson, Lisa E. “R740S Mutation of Tcirg1 Mutation Affects Enamel Development in Osteopetrotic Mice.” 2016. Masters Thesis, University of Toronto. Accessed April 17, 2021.
http://hdl.handle.net/1807/74943.
MLA Handbook (7th Edition):
Johnson, Lisa E. “R740S Mutation of Tcirg1 Mutation Affects Enamel Development in Osteopetrotic Mice.” 2016. Web. 17 Apr 2021.
Vancouver:
Johnson LE. R740S Mutation of Tcirg1 Mutation Affects Enamel Development in Osteopetrotic Mice. [Internet] [Masters thesis]. University of Toronto; 2016. [cited 2021 Apr 17].
Available from: http://hdl.handle.net/1807/74943.
Council of Science Editors:
Johnson LE. R740S Mutation of Tcirg1 Mutation Affects Enamel Development in Osteopetrotic Mice. [Masters Thesis]. University of Toronto; 2016. Available from: http://hdl.handle.net/1807/74943
University of Toronto
22.
Saw, Ner Mu Nar.
The Roles of the Voa Subunit of the Vacuolar H+-ATPase in Dense-core Vesicle Acidification, Transmitter Uptake and Storage.
Degree: 2011, University of Toronto
URL: http://hdl.handle.net/1807/31433
► The Vo sector of the vacuolar H+-ATPase is a multi-subunit complex that forms a proteolipid pore. The largest subunit in this complex is the a…
(more)
▼ The Vo sector of the vacuolar H+-ATPase is a multi-subunit complex that forms a proteolipid pore. The largest subunit in this complex is the a subunit which has four isoforms (a1-a4). The isoform(s) critical for secretory vesicle acidification has yet to be identified. Using a cell line derived from rat pheochromocytoma in which Voa1 and/or Voa2 had been down-regulated this study revealed that Voa1, and to a lesser extent, Voa2 are critical for acidifying dense-core vesicles (DCVs). The acidification defects resulting from down-regulation of Voa1 and Voa1/ Voa2 were suppressed by the expression of knockdown-resistant Voa1. Defects in DCV acidification resulted in reductions in their transmitter uptake and storage. Lastly, Ca2+-dependent peptide secretion appeared normal in Voa1 and Voa1/ Voa2 knockdown cells. . This study demonstrated that Voa1 and Voa2 cooperatively regulate dense-core vesicle acidification as well as transmitter uptake/storage, while they may not be critical for dense-core vesicle exocytosis.
MAST
Advisors/Committee Members: Sugita, Shuzo, Physiology.
Subjects/Keywords: V-ATPase; Voa; transmitter uptake; acidification; 0719
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Manager
APA (6th Edition):
Saw, N. M. N. (2011). The Roles of the Voa Subunit of the Vacuolar H+-ATPase in Dense-core Vesicle Acidification, Transmitter Uptake and Storage. (Masters Thesis). University of Toronto. Retrieved from http://hdl.handle.net/1807/31433
Chicago Manual of Style (16th Edition):
Saw, Ner Mu Nar. “The Roles of the Voa Subunit of the Vacuolar H+-ATPase in Dense-core Vesicle Acidification, Transmitter Uptake and Storage.” 2011. Masters Thesis, University of Toronto. Accessed April 17, 2021.
http://hdl.handle.net/1807/31433.
MLA Handbook (7th Edition):
Saw, Ner Mu Nar. “The Roles of the Voa Subunit of the Vacuolar H+-ATPase in Dense-core Vesicle Acidification, Transmitter Uptake and Storage.” 2011. Web. 17 Apr 2021.
Vancouver:
Saw NMN. The Roles of the Voa Subunit of the Vacuolar H+-ATPase in Dense-core Vesicle Acidification, Transmitter Uptake and Storage. [Internet] [Masters thesis]. University of Toronto; 2011. [cited 2021 Apr 17].
Available from: http://hdl.handle.net/1807/31433.
Council of Science Editors:
Saw NMN. The Roles of the Voa Subunit of the Vacuolar H+-ATPase in Dense-core Vesicle Acidification, Transmitter Uptake and Storage. [Masters Thesis]. University of Toronto; 2011. Available from: http://hdl.handle.net/1807/31433
University of Texas Southwestern Medical Center
23.
Sauer, David Bryant.
Protein Structure and Ion Binding in Potassium Selective Channels.
Degree: 2013, University of Texas Southwestern Medical Center
URL: http://hdl.handle.net/2152.5/1262
► Potassium channels play a central role in a number of biological processes, most classically the action potential of excitable cells in multicellular organisms. These channels…
(more)
▼ Potassium channels play a central role in a number of biological processes, most classically the action potential of excitable cells in multicellular organisms. These channels are defined by their selective conduction of potassium to the exclusion of other monovalent ions as governed by a common sequence and structural motif, the selectivity filter. This structure, made of backbone carbonyls and threonine side chains, directly coordinates the ions as they diffuse through the channel and appears central to this sub-angstrom discrimination between cations. Utilizing the non-selective Sodium and Potassium conducting channel (NaK) as a structural scaffold the mechanisms of both ion selectivity and formation of this selective structure are examined.
Advisors/Committee Members: Jiang, Youxing.
Subjects/Keywords: Sodium-Potassium-Exchanging ATPase; Potassium; Models, Molecular
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APA (6th Edition):
Sauer, D. B. (2013). Protein Structure and Ion Binding in Potassium Selective Channels. (Thesis). University of Texas Southwestern Medical Center. Retrieved from http://hdl.handle.net/2152.5/1262
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Chicago Manual of Style (16th Edition):
Sauer, David Bryant. “Protein Structure and Ion Binding in Potassium Selective Channels.” 2013. Thesis, University of Texas Southwestern Medical Center. Accessed April 17, 2021.
http://hdl.handle.net/2152.5/1262.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Sauer, David Bryant. “Protein Structure and Ion Binding in Potassium Selective Channels.” 2013. Web. 17 Apr 2021.
Vancouver:
Sauer DB. Protein Structure and Ion Binding in Potassium Selective Channels. [Internet] [Thesis]. University of Texas Southwestern Medical Center; 2013. [cited 2021 Apr 17].
Available from: http://hdl.handle.net/2152.5/1262.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Sauer DB. Protein Structure and Ion Binding in Potassium Selective Channels. [Thesis]. University of Texas Southwestern Medical Center; 2013. Available from: http://hdl.handle.net/2152.5/1262
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Colorado State University
24.
Tapken, Wiebke.
Copper transport into the chloroplast and its implications for copper homeostasis in Arabidopsis thaliana.
Degree: PhD, Biology, 2012, Colorado State University
URL: http://hdl.handle.net/10217/71670
► Copper (Cu) is an essential micronutrient for most aerobic organisms including plants. It is present as Cu+ or Cu2+, which makes it an ideal cofactor…
(more)
▼ Copper (Cu) is an essential micronutrient for most aerobic organisms including plants. It is present as Cu+ or Cu2+, which makes it an ideal cofactor for enzymes involved in processes such as photosynthesis and respiration. Plant cuproproteins are almost ubiquitously found in every cell compartment. The blue Cu protein plastocyanin (PC) is believed to bind the majority of Cu ions in green tissues and is essential for higher plants. Cu reaches the thylakoid lumen through the activity of two P1B-type ATPases called PAA1/HMA6 and PAA2/HMA8 (P-type
ATPase of Arabidopsis/Heavy-metal
ATPase), which are located in the inner chloroplast envelope and the thylakoid lumen respectively. Under Cu limiting conditions, plants have been suggested to prioritize cellular Cu to PC to ensure adequate photosynthesis. This process involves the post-transcriptional down-regulation of seemingly less essential cuproproteins through the activity of a single transcription factor called SPL7 (SQUAMOSA promoter binding protein-like7). The first chapter reviews Cu homeostasis in plants. The research presented in the three experimental chapters of this dissertation is aimed to determine the role of the chloroplast in Cu homeostasis of Arabidopsis thaliana. I report a novel SPL7-independent and chloroplast-specific regulation of the thylakoid-localized Cu transporter PAA2/HMA8. The transporter is most abundant in the absence of Cu and is turned over at higher chloroplastic Cu concentrations. PAA2/HMA8 abundance in Cu deficiency is furthermore controlled by the presence of PC, because in a pc mutant PAA2/HMA8 abundance is always low. The regulation of the transporter likely serves as a checkpoint for the Cu requirements of the thylakoid lumen. I identified two components of the stroma-localized Clp protease (Caseinolytic peptidase) which are involved in PAA2/HMA8 turnover. The Cu status of these mutants is not affected, decreasing the likelihood of a secondary affect of Cu on PAA2/HMA8 in these plants. In the last experimental chapter I summarize relevant results that further describe and characterize PAA1 and PAA2. Most notably, Arabidopsis encodes for a splice-form of PAA1. This much smaller fragment is expressed with a chloroplast targeting sequence and could potentially function as a stromal Cu chaperone.
Advisors/Committee Members: Pilon, Marinus (advisor), Chisholm, Stephen (committee member), Pilon-Smits, Elizabeth (committee member), Reddy, Anireddy S.N. (committee member).
Subjects/Keywords: homeostasis; copper; P type ATPase; chloroplast; Arabidopsis
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APA (6th Edition):
Tapken, W. (2012). Copper transport into the chloroplast and its implications for copper homeostasis in Arabidopsis thaliana. (Doctoral Dissertation). Colorado State University. Retrieved from http://hdl.handle.net/10217/71670
Chicago Manual of Style (16th Edition):
Tapken, Wiebke. “Copper transport into the chloroplast and its implications for copper homeostasis in Arabidopsis thaliana.” 2012. Doctoral Dissertation, Colorado State University. Accessed April 17, 2021.
http://hdl.handle.net/10217/71670.
MLA Handbook (7th Edition):
Tapken, Wiebke. “Copper transport into the chloroplast and its implications for copper homeostasis in Arabidopsis thaliana.” 2012. Web. 17 Apr 2021.
Vancouver:
Tapken W. Copper transport into the chloroplast and its implications for copper homeostasis in Arabidopsis thaliana. [Internet] [Doctoral dissertation]. Colorado State University; 2012. [cited 2021 Apr 17].
Available from: http://hdl.handle.net/10217/71670.
Council of Science Editors:
Tapken W. Copper transport into the chloroplast and its implications for copper homeostasis in Arabidopsis thaliana. [Doctoral Dissertation]. Colorado State University; 2012. Available from: http://hdl.handle.net/10217/71670
Univerzitet u Beogradu
25.
Jovanović, Aleksandra A., 1987- 15120743.
Signalni putevi estradiola uključeni u regulaciju
ekspresije i aktivnosti inducibilne azot-monoksid-sintaze i
natrijum-kalijum adenozintrifosfataze u srcu gojaznih ženki
pacova.
Degree: Biološki fakultet, 2019, Univerzitet u Beogradu
URL: https://fedorabg.bg.ac.rs/fedora/get/o:19580/bdef:Content/get
► Biologija - Molekularna endokrinologija / Biology - Molecular Endocrinology
Gojaznost je oboljenje povezano sa nizom patoloških stanja kao što su: rezistencija na insulin (IR), kardiovaskularne…
(more)
▼ Biologija - Molekularna endokrinologija / Biology -
Molecular Endocrinology
Gojaznost je oboljenje povezano sa nizom patoloških
stanja kao što su: rezistencija na insulin (IR), kardiovaskularne
bolesti (KVB) i Diabetes Mellitus tipa 2 (DMT2). Povećana
ekspresija i aktivnost inducibilne azot-monoksid-sintaze (iNOS;
engl. Inducible Nitric Oxide Synthase) u srcu u stanju gojaznosti,
moţe dovesti do apoptoze kardiomiocita i hipertrofije srca, dok sa
druge strane gojaznost zdruţena sa IR doprinosi smanjenoj
aktivnosti Na+/K+-ATPaze, što dovodi do smanjenja kontraktilnosti
vaskulature i razvoja sistemske hipertenzije. Endogeni estradiol
svojim delovanjem sprečava nastanak IR i hiperglikemije i ostvaruje
pozitivne efekte na kardiovaskularni sistem (KVS), ali sinteza i
kardioprotektivni uticaj estradiola mogu biti smanjeni usled
razvoja gojaznosti. Estradiol ostvaruje kardioprotektivne tako što
utiče na smanjenje ekspresije i aktivnosti iNOS, kao i povećanje
ekspresije i aktivnosti Na+/K+-ATPaze, posredstvom različitih
signalnih molekula i kinaza, kao što su: supstrat receptora za
insulin 1 (IRS-1; engl. Insulin Receptor Substrate)/
fosfatidil-inozitol-3-kinaza (PI3K; engl. Phosphatidylinositide
3-Kinase)/ protein kinaza B (Akt; engl. Protein Kinase B),
ekstracelularnim signalima regulisane kinaze 1 i 2 (ERK1/2; engl.
Extracellular Signal-Regulated Protein Kinases 1 i 2) kao i RhoA
(engl. Ras homolog gene family, member A)/ROCK (engl.
Rho-associated protein kinase). Pored direktnog efekta na srce,
estradiol posredno reguliše i njegovu funkciju tako sto utiče na
metabolizam i transport glukoze i SMK, preko transportera glukoze
(GLUT; engl. Glucose Transporters) i translokaze masnih kiselina
(FAT; CD36 ; engl. Fatty Acid Translocase). Za izradu ove doktorske
disertacije je korišćeno 16 adultnih ţenki pacova soja Wistar,
podeljenih u dve eksperimentalne grupe. Prva grupa pacova je tokom
10 nedelja hranjena standardnom laboratorijskom hranom za pacove,
dok je druga grupa pacova tokom 10 nedelja hranjena standardnom
laboratorijskom hranom obogaćenom sa 42% masti (HF reţim ishrane).
Nakon 10 nedelja pacovi su ţrtvovani, sakupljena je krv i izolovan
je serum, a srca su ekstrahovana i delovi tkiva su korišćeni za
izolovanje proteina i RNK. U serumu pacova je odreĎivana
koncentracija estradiola, dok su u lizatu srca pacova odreĎivane
koncentracije L-Arginina (L-Arg), NO i slobodnih masnih kiselina
(SMK). Metodom qRT-PCR odreĎivan je nivo iRNK iNOS u srcu
pacova...
Advisors/Committee Members: Isenović, Esma, 1962- 3563367.
Subjects/Keywords: iNOS; Na+/K+-ATPase; obesity; estradiol;
heart
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Manager
APA (6th Edition):
Jovanović, Aleksandra A., 1. 1. (2019). Signalni putevi estradiola uključeni u regulaciju
ekspresije i aktivnosti inducibilne azot-monoksid-sintaze i
natrijum-kalijum adenozintrifosfataze u srcu gojaznih ženki
pacova. (Thesis). Univerzitet u Beogradu. Retrieved from https://fedorabg.bg.ac.rs/fedora/get/o:19580/bdef:Content/get
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Chicago Manual of Style (16th Edition):
Jovanović, Aleksandra A., 1987- 15120743. “Signalni putevi estradiola uključeni u regulaciju
ekspresije i aktivnosti inducibilne azot-monoksid-sintaze i
natrijum-kalijum adenozintrifosfataze u srcu gojaznih ženki
pacova.” 2019. Thesis, Univerzitet u Beogradu. Accessed April 17, 2021.
https://fedorabg.bg.ac.rs/fedora/get/o:19580/bdef:Content/get.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Jovanović, Aleksandra A., 1987- 15120743. “Signalni putevi estradiola uključeni u regulaciju
ekspresije i aktivnosti inducibilne azot-monoksid-sintaze i
natrijum-kalijum adenozintrifosfataze u srcu gojaznih ženki
pacova.” 2019. Web. 17 Apr 2021.
Vancouver:
Jovanović, Aleksandra A. 11. Signalni putevi estradiola uključeni u regulaciju
ekspresije i aktivnosti inducibilne azot-monoksid-sintaze i
natrijum-kalijum adenozintrifosfataze u srcu gojaznih ženki
pacova. [Internet] [Thesis]. Univerzitet u Beogradu; 2019. [cited 2021 Apr 17].
Available from: https://fedorabg.bg.ac.rs/fedora/get/o:19580/bdef:Content/get.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Jovanović, Aleksandra A. 11. Signalni putevi estradiola uključeni u regulaciju
ekspresije i aktivnosti inducibilne azot-monoksid-sintaze i
natrijum-kalijum adenozintrifosfataze u srcu gojaznih ženki
pacova. [Thesis]. Univerzitet u Beogradu; 2019. Available from: https://fedorabg.bg.ac.rs/fedora/get/o:19580/bdef:Content/get
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
University of Melbourne
26.
RIEDMAIER, PATRICE.
Microbial NTPDases and their contribution to infection.
Degree: 2012, University of Melbourne
URL: http://hdl.handle.net/11343/37790
► Legionella pneumophila is an intracellular bacterium that has adapted to infect human hosts. Infection of susceptible individuals can result in the onset of severe atypical…
(more)
▼ Legionella pneumophila is an intracellular bacterium that has adapted to infect human hosts. Infection of susceptible individuals can result in the onset of severe atypical pneumonia commonly referred to as Legionnaires’ disease. Genome sequence analysis of L. pneumophila has revealed the presence of numerous eukaryotic-like proteins and proteins with eukaryotic domains. These include two apyrases annotated lpg0971 and lpg1905, which are homologues of human CD39/NTPDase1. Lpg0971 and Lpg1905 belong to the GDA1_CD39 superfamily that are characterised by their ability to hydrolyse terminal phosphoanhydride bonds of extracellular nucleoside di- and tri-phosphates, such as ATP and ADP. Prior to the commencement of this study, the importance of Lpg1905 to bacterial infection has been established. In this study, the contribution of both NTPDases to Legionella infection was investigated, in particular the role of Lpg0971 during infection and further structural and functional characterisation of Lpg1905.
A biochemical assay that measures the release of inorganic phosphate was used to determine the functional characteristics of recombinant, refolded Lpg0971. Highest enzymatic function was observed in the presence of manganese (II) ions. Additionally, Lpg0971 displayed affinity for ATP hydrolysis only. To ascertain whether Lpg0971 contributed to vacuolar replication within host cells, bacterial replication was tested in human alveolar macrophages and amoebae. The lpg0971 mutant was unable to replicate over 72 h in both cell lines; however, complementation of the mutant restored replication. Moreover, the two NTPDases appeared to be functionally redundant during replication in macrophages.
The elucidation of the crystal structure of Lpg1905 during the course of this study provided insight into the residues surrounding the substrate in the active site. Three residues were selected for their proximity to the substrate binding pocket and targeted site-directed mutagenesis was performed. These studies revealed the importance of Arg56 for binding the substrate during hydrolysis and Try346 for positioning the substrate during hydrolysis. The conservative mutation R56K resulted in a mutant derivative with NTPase activity only (35% of wild type activity), whereas the nonconservative mutation Y346A produced a mutant derivative with higher NDPase activity (35% of wild type activity) than NTPase activity (20% of wild type activity). Comparison of the ability of these mutant derivatives to complement lpg1905 mutant during in vitro replication assays within THP-1 macrophages demonstrated the importance of NTPase activity rather than NDPase activity of Lpg1905 to bacterial replication. This was supported by a chimeric mutant derivative in which the ACR1 motif of Lpg1905 was mutated from TGSR to SHTS that exhibited only NTPase activity (40% of wild type activity) but was also able to replicate within THP-1 macrophages. Lastly, enzyme kinetic comparisons…
Subjects/Keywords: pathogenesis; bacterial; legionella pneumophila; NTPDase; ATPase; inhibitors
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APA (6th Edition):
RIEDMAIER, P. (2012). Microbial NTPDases and their contribution to infection. (Doctoral Dissertation). University of Melbourne. Retrieved from http://hdl.handle.net/11343/37790
Chicago Manual of Style (16th Edition):
RIEDMAIER, PATRICE. “Microbial NTPDases and their contribution to infection.” 2012. Doctoral Dissertation, University of Melbourne. Accessed April 17, 2021.
http://hdl.handle.net/11343/37790.
MLA Handbook (7th Edition):
RIEDMAIER, PATRICE. “Microbial NTPDases and their contribution to infection.” 2012. Web. 17 Apr 2021.
Vancouver:
RIEDMAIER P. Microbial NTPDases and their contribution to infection. [Internet] [Doctoral dissertation]. University of Melbourne; 2012. [cited 2021 Apr 17].
Available from: http://hdl.handle.net/11343/37790.
Council of Science Editors:
RIEDMAIER P. Microbial NTPDases and their contribution to infection. [Doctoral Dissertation]. University of Melbourne; 2012. Available from: http://hdl.handle.net/11343/37790
Colorado State University
27.
Springer, Courtney Lee.
Characterization of poliovirus 2CATPase bound to bilayer nanodiscs and involvement of the poliovirus 3Dpol thumb α-helix in determining poly(A) tail length.
Degree: PhD, Biochemistry and Molecular Biology, 2013, Colorado State University
URL: http://hdl.handle.net/10217/80976
► Poliovirus (PV) is a small non-enveloped picornavirus with a ≈7.5 kb long single-stranded, positive-sense RNA genome. Upon infection, the RNA is translated to generate a…
(more)
▼ Poliovirus (PV) is a small non-enveloped picornavirus with a ≈7.5 kb long single-stranded, positive-sense RNA genome. Upon infection, the RNA is translated to generate a ≈250 kDa polyprotein that is subsequently cleaved into about a dozen fully processed proteins and several functional intermediates. PV replication occurs in large membrane associated complexes involving the "non-structural" P2 and P3 region proteins and two of these proteins, 2CATPase and 3Dpol, are the subjects of this dissertation. Part I of this work is focused on the 2C protein, an AAA+ family
ATPase that plays a key role in host cell membrane rearrangements and virion assembly, but the membrane binding characteristics of 2C and its polyprotein precursors have made it difficult to elucidate their exact roles in virus replication. In this work I show that small lipid bilayers known as nanodiscs can be used to chaperone the in vitro expression of soluble poliovirus 2C and the precursor 2BC and 2BC3AB polyproteins in a membrane bound form. Biochemical analysis shows that the proteins are highly active over a wide range of salt concentrations, exhibit slight lipid headgroup dependence, and show significant stimulation by acetate. Notably, the
ATPase activity of the core 2C domain is stimulated ≈60-fold as compared to the larger 2BC3AB polyprotein, with most of this stimulation occurring upon removal of 2B. This data leads to a model wherein the viral replication complex can be assembled with a minimally active form of 2C that then becomes fully activated upon proteolytic cleavage from the adjacent 2B viroporin domain. In Part II of this dissertation, I focus on the role of the viral RNA polymerase, 3Dpol, in maintaining the ≈20-150 nucleotides long 3' poly(A) tail of the viral genome. The length of the tail is important for viral replication and initiation of (-)-strand synthesis, but the means by which the RNA is polyadenylated and how poly(A) tail length is regulated is not well understood. We have identified several mutations in an α-helix of the 3Dpol thumb domain that directly impact poly(A) tail length. Here, I tested the impact of these mutations on reiterative transcription of poly(A), poly(U), and poly(C) templates as well as characterized their effect on 3Dpol initiation, stability, elongation rate, and fidelity. I found that mutations in the thumb have the greatest impact on elongation complex stability and that 3Dpol is able to reiteratively transcribe homopolymeric poly(U) and poly(A), but not poly(C) RNA templates. Interestingly, distinct poly(A) and poly(U) transcripts are generated from 10 nucleotide homopolymers that are 1, 7, or 8 nucleotides longer than the template. Based on these findings, we propose a poly(A) slippage model in which the elongation complex stalls at the end of the homopolymer stretch in the absence of additional nucleotides to promote a single nucleotide slippage. This is followed by a slow structural rearrangement in which 3Dpol slips back to the 3' end of the homopolymer sequence, where it is…
Advisors/Committee Members: Peersen, Olve B. (advisor), Ho, P. Shing (committee member), Luger, Karolin (committee member), Kennan, Alan (committee member).
Subjects/Keywords: enzymology; membrane protein; poliovirus; polymerase; virology; ATPase
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APA (6th Edition):
Springer, C. L. (2013). Characterization of poliovirus 2CATPase bound to bilayer nanodiscs and involvement of the poliovirus 3Dpol thumb α-helix in determining poly(A) tail length. (Doctoral Dissertation). Colorado State University. Retrieved from http://hdl.handle.net/10217/80976
Chicago Manual of Style (16th Edition):
Springer, Courtney Lee. “Characterization of poliovirus 2CATPase bound to bilayer nanodiscs and involvement of the poliovirus 3Dpol thumb α-helix in determining poly(A) tail length.” 2013. Doctoral Dissertation, Colorado State University. Accessed April 17, 2021.
http://hdl.handle.net/10217/80976.
MLA Handbook (7th Edition):
Springer, Courtney Lee. “Characterization of poliovirus 2CATPase bound to bilayer nanodiscs and involvement of the poliovirus 3Dpol thumb α-helix in determining poly(A) tail length.” 2013. Web. 17 Apr 2021.
Vancouver:
Springer CL. Characterization of poliovirus 2CATPase bound to bilayer nanodiscs and involvement of the poliovirus 3Dpol thumb α-helix in determining poly(A) tail length. [Internet] [Doctoral dissertation]. Colorado State University; 2013. [cited 2021 Apr 17].
Available from: http://hdl.handle.net/10217/80976.
Council of Science Editors:
Springer CL. Characterization of poliovirus 2CATPase bound to bilayer nanodiscs and involvement of the poliovirus 3Dpol thumb α-helix in determining poly(A) tail length. [Doctoral Dissertation]. Colorado State University; 2013. Available from: http://hdl.handle.net/10217/80976
University of Kansas
28.
Jansson, Kyle.
Novel effects of ouabain in autosomal dominant polycystic kidney disease cystogenesis.
Degree: PhD, Molecular & Integrative Physiology, 2013, University of Kansas
URL: http://hdl.handle.net/1808/19625
► Autosomal dominant polycystic kidney disease (ADPKD) is caused by mutations in Pkd1 or Pkd2, genes encoding for polycystin-1 (PC-1) and polycystin-2 (PC-2), respectively. ADPKD is…
(more)
▼ Autosomal dominant polycystic kidney disease (ADPKD) is caused by mutations in Pkd1 or Pkd2, genes encoding for polycystin-1 (PC-1) and polycystin-2 (PC-2), respectively. ADPKD is characterized by the progressive growth of numerous fluid-filled renal cysts. Cyst formation and growth depends on proliferation of the cyst-lining epithelial cells and fluid secretion into the cyst lumen. ADPKD cystogenesis is highly influenced by non-genomic factors, many of which elicit their effects via cAMP-dependent pathways. Understanding mechanisms mediating the effects of cystogenic agents is crucial for the future development of ADPKD therapy. Previous work has shown that cells derived from the epithelial-lining of renal cysts from patients with ADPKD (ADPKD cells) have an increased affinity for the hormone ouabain. ADPKD cells respond to ouabain by an increased rate of cell proliferation. This effect depends on binding of ouabain to the Na,K-
ATPase which induces activation of Src kinase, epidermal growth factor receptor (EGFR), and the extracellular regulated kinase (ERK1/2) pathway. The objective of the current study was to determine the role of ouabain in mechanisms of fluid secretion and cyst growth in ADPKD. Studies were carried out in human ADPKD cells, embryonic kidneys from the Pkd1m1Bei mouse model, and M-1 mouse cortical collecting duct cells. Results of this study show that physiologic concentrations of ouabain enhance cAMP-dependent fluid secretion and cyst growth of ADPKD cells grown in culture as monolayers or in three-dimensional structures resembling cysts. Additionally, ouabain potentiated the cAMP-dependent growth of cyst-like dilations in metanephric kidneys from the Pkd1m1Bei mouse model. These effects were mediated via activation of the Na,K-
ATPase signaling apparatus, located at the basolateral domain of ADPKD cells. Intracellular mediators of ouabain's response included the EGFR-Src-ERK pathway. Ouabain alone did not increase fluid secretion and cyst growth. Rather, ouabain treatment altered the phenotype of ADPKD cells to allow enhanced responses to cAMP agonists. The potentiating effect of ouabain on cAMP-induced fluid secretion was associated with the capacity of ouabain to stimulate anion secretion via the apically located cystic fibrosis transmembrane conductance regulator (CFTR). Moreover, ouabain increased membrane expression of the CFTR. Finally, ouabain decreased Na,K-
ATPase membrane expression and ion transport at the basolateral membrane of ADPKD cells. The increased ouabain sensitivity of ADPKD cells depends on an abnormally high affinity of the Na,K-
ATPase for ouabain. Increased ouabain affinity of the Na,K-
ATPase was associated with abnormal expression of the C-terminus of PC-1 in M-1 cells. Altogether, the study of ouabain's effects in ADPKD have uncovered a novel role for ouabain as a physiologic agent that influences renal cyst growth in ADPKD. In addition, it has identified a new mechanism in ADPKD cystogenesis, important for the progression of the disease.
Advisors/Committee Members: Blanco, Gustavo (advisor), Fields, Timothy A (cmtemember), Grantham, Jared J (cmtemember), Wolfe, Michael W (cmtemember), Wood, John G (cmtemember).
Subjects/Keywords: Physiology; Medicine; ADPKD; Na; K-ATPase; ouabain
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APA (6th Edition):
Jansson, K. (2013). Novel effects of ouabain in autosomal dominant polycystic kidney disease cystogenesis. (Doctoral Dissertation). University of Kansas. Retrieved from http://hdl.handle.net/1808/19625
Chicago Manual of Style (16th Edition):
Jansson, Kyle. “Novel effects of ouabain in autosomal dominant polycystic kidney disease cystogenesis.” 2013. Doctoral Dissertation, University of Kansas. Accessed April 17, 2021.
http://hdl.handle.net/1808/19625.
MLA Handbook (7th Edition):
Jansson, Kyle. “Novel effects of ouabain in autosomal dominant polycystic kidney disease cystogenesis.” 2013. Web. 17 Apr 2021.
Vancouver:
Jansson K. Novel effects of ouabain in autosomal dominant polycystic kidney disease cystogenesis. [Internet] [Doctoral dissertation]. University of Kansas; 2013. [cited 2021 Apr 17].
Available from: http://hdl.handle.net/1808/19625.
Council of Science Editors:
Jansson K. Novel effects of ouabain in autosomal dominant polycystic kidney disease cystogenesis. [Doctoral Dissertation]. University of Kansas; 2013. Available from: http://hdl.handle.net/1808/19625
29.
DeLaney, Elizabeth Erin.
RNA Recognition by the Pattern Recognition Receptor RIG-I:
Roles of RNA Binding, Multimerization, and RNA-dependent ATPase
Activity.
Degree: PhD, Biochemistry, 2014, Case Western Reserve University School of Graduate Studies
URL: http://rave.ohiolink.edu/etdc/view?acc_num=case1405015903
► Recognition of viral RNA by mammalian cells is critical for the activation of the innate immune system. Viral RNA is recognized by several pathogen recognition…
(more)
▼ Recognition of viral RNA by mammalian cells is
critical for the activation of the innate immune system. Viral RNA
is recognized by several pathogen recognition receptors, including
retinoic acid inducible gene I, or RIG-I. RIG-I consists of two
N-terminal tandem caspase activation and recruitment domains, a
central helicase/
ATPase domain, and a C-terminal regulatory domain.
Following RNA binding, RIG-I undergoes a conformational change,
ubiquitination, and dimerization, all of which are necessary for
interaction with the adaptor protein mitochondrial antiviral
signaling (MAVS). Binding to MAVS triggers signaling cascades that
induce the transcription of antiviral peptides. RIG-I has been
shown to be activated by both dsRNA and dsRNA containing
5’-triphosphates in vivo, and its
ATPase activity is critical for
activation. A significant body of work has been published regarding
the cellular role of RIG-I, but how RIG-I distinguishes viral RNAs
from cellular RNAs remains unclear. To understand how RIG-I
distinguishes between different substrates, we performed a
biochemical analysis of RIG-I RNA binding,
ATPase activity, and
oligomerization. We used purified RIG-I to quantitatively analyze
how RIG-I interacts with various model RNAs. We show that RIG-I
binds tightly to dsRNA regardless of the presence of a 5’-
triphosphate. Dissociation of RIG-I from RNA is enhanced by ATP.
RIG-I
ATPase activity is stimulated by RNA duplexes as short as 10
bp, and a RIG-I monomer is sufficient for
ATPase activity. RIG-I
binds to RNA duplexes with and without blunt ends, however
ATPase
activity is only activated by RNA duplexes containing at least one
blunt end. Collectively, these data suggest that duplex structure
and nucleotide binding play a critical role in RIG-I binding and
activation. Our data suggest a model in which distinguishing self
from non-self RNA requires the recognition of multiple features in
a single RNA by RIG-I.
Advisors/Committee Members: Jankowsky, Eckhard (Advisor).
Subjects/Keywords: Biochemistry; Innate immunity; RIG-I; ATPase
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APA (6th Edition):
DeLaney, E. E. (2014). RNA Recognition by the Pattern Recognition Receptor RIG-I:
Roles of RNA Binding, Multimerization, and RNA-dependent ATPase
Activity. (Doctoral Dissertation). Case Western Reserve University School of Graduate Studies. Retrieved from http://rave.ohiolink.edu/etdc/view?acc_num=case1405015903
Chicago Manual of Style (16th Edition):
DeLaney, Elizabeth Erin. “RNA Recognition by the Pattern Recognition Receptor RIG-I:
Roles of RNA Binding, Multimerization, and RNA-dependent ATPase
Activity.” 2014. Doctoral Dissertation, Case Western Reserve University School of Graduate Studies. Accessed April 17, 2021.
http://rave.ohiolink.edu/etdc/view?acc_num=case1405015903.
MLA Handbook (7th Edition):
DeLaney, Elizabeth Erin. “RNA Recognition by the Pattern Recognition Receptor RIG-I:
Roles of RNA Binding, Multimerization, and RNA-dependent ATPase
Activity.” 2014. Web. 17 Apr 2021.
Vancouver:
DeLaney EE. RNA Recognition by the Pattern Recognition Receptor RIG-I:
Roles of RNA Binding, Multimerization, and RNA-dependent ATPase
Activity. [Internet] [Doctoral dissertation]. Case Western Reserve University School of Graduate Studies; 2014. [cited 2021 Apr 17].
Available from: http://rave.ohiolink.edu/etdc/view?acc_num=case1405015903.
Council of Science Editors:
DeLaney EE. RNA Recognition by the Pattern Recognition Receptor RIG-I:
Roles of RNA Binding, Multimerization, and RNA-dependent ATPase
Activity. [Doctoral Dissertation]. Case Western Reserve University School of Graduate Studies; 2014. Available from: http://rave.ohiolink.edu/etdc/view?acc_num=case1405015903
University of Toronto
30.
Ripstein, Zev Aaron.
Structural Insights into AAA+ Unfoldases and Associated Proteases.
Degree: PhD, 2020, University of Toronto
URL: http://hdl.handle.net/1807/103627
► Ring-shaped unfoldases of the AAA+ (ATPases Associated with a variety of cellular Activities) su- perfamily are responsible for the dismantling or remodelling of various protein…
(more)
▼ Ring-shaped unfoldases of the AAA+ (ATPases Associated with a variety of cellular Activities) su- perfamily are responsible for the dismantling or remodelling of various protein complexes in the cell. These hexameric enzymes utilize energy from ATP hydrolysis to unfold client proteins, often passing them to proteases to be recycled into their amino acid components. Investigating the interplay between AAA+ motors, proteases, and substrates is key to understanding protein homeostasis in the cell. To investigate these systems, a combined structural approach using single particle electron cryomicroscopy (cryo-EM) and Nuclear Magnetic Resonance (NMR) spectroscopy was taken. Utilizing these approaches enabled structural insights into the archaeal Cdc48 system from Thermoplasma acidophilum, as well as the bacterial ClpP systems from Staphyloccoccus aureus, Mycobacterium tuberculosis, and Neisseria meningitidis. Our results illustrate several mechanisms of allosteric control, as well as protein-protein interactions that are important for proper function.
Advisors/Committee Members: Rubinstein, John L, Kay, Lewis E, Biochemistry.
Subjects/Keywords: ATPase; CryoEM; NMR; Protease; Protein; Unfolding; 0487
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APA ·
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APA (6th Edition):
Ripstein, Z. A. (2020). Structural Insights into AAA+ Unfoldases and Associated Proteases. (Doctoral Dissertation). University of Toronto. Retrieved from http://hdl.handle.net/1807/103627
Chicago Manual of Style (16th Edition):
Ripstein, Zev Aaron. “Structural Insights into AAA+ Unfoldases and Associated Proteases.” 2020. Doctoral Dissertation, University of Toronto. Accessed April 17, 2021.
http://hdl.handle.net/1807/103627.
MLA Handbook (7th Edition):
Ripstein, Zev Aaron. “Structural Insights into AAA+ Unfoldases and Associated Proteases.” 2020. Web. 17 Apr 2021.
Vancouver:
Ripstein ZA. Structural Insights into AAA+ Unfoldases and Associated Proteases. [Internet] [Doctoral dissertation]. University of Toronto; 2020. [cited 2021 Apr 17].
Available from: http://hdl.handle.net/1807/103627.
Council of Science Editors:
Ripstein ZA. Structural Insights into AAA+ Unfoldases and Associated Proteases. [Doctoral Dissertation]. University of Toronto; 2020. Available from: http://hdl.handle.net/1807/103627
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Open Access Theses and Dissertations
