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University of Waikato
1.
Cumming, Mathew Hoani.
Structural and enzymatic characterisation of nucleoside triphosphate diphosphohydrolases from Trifolium repens and Dolichos biflorus
.
Degree: 2012, University of Waikato
URL: http://hdl.handle.net/10289/6331 
► Extracellular nucleoside triphosphate diphosphohydrolases (NTPDases) are enzymes that reduce the extracellular nucleotide signal and inactivate the purinogenic signalling pathway. These enzymes, in the presence of…
(more)
▼ Extracellular nucleoside triphosphate diphosphohydrolases (NTPDases) are enzymes that reduce the extracellular nucleotide signal and inactivate the purinogenic signalling pathway. These enzymes, in the presence of a divalent cation sequentially hydrolyses the γ- and β- phosphoanhydride bond from a range of nucleotide di- or triphosphates to the corresponding nucleotide monophosphate.
There is evidence that purinogenic signalling is present in higher plants and it is becoming clear that NTPDases play a role in the early stages of rhizobium infection during nodulation in legumes. To further our understanding of NTPDases, this study investigated the biochemical and structural characteristics of two legume NTPDases believed to be involved in nodulation. The first, 7WC, was isolated from the roots of white clover and the second, DbLnP, from the roots of Dolichos biflorus. DbLnP has been characterised as a carbohydrate binding NTPDase that is directly associated with the perception of rhizobia. Using X-ray crystallography a number of crystal structures of 7WC and DbLnP were determined at resolutions between 1.9 Å and 2.9 Å. For 7WC, structures were determined for an apo- form, an AMP-bound and also bound with the nonhydrolysable
ATP analogue AMPPNP. For DbLnP structures were solved with phosphate and Mn2+ bound and another with AMPPNP and Mn2+ bound. Kinetic analysis of a range of substrates together with the analysis of the binding modes of 7WC and DbLnP explained substrate preference for each of the NTPDases.
These analyses showed that NTPDases can adopt two conformations depending on substrate and co-factor binding. The central hinge region creates a ‘butterfly’ motion of the domains that reduces the width of the active site cleft. This phenomenon has been previously hypothesised but has not been observed for NTPDases.
A model of catalysis is proposed whereby the ‘open’ form first binds substrate in an inactive orientation. Binding of the metal ion induces a conformational change that brings the domains together and allows movement of the phosphate tail deeper into the active site cleft – a reorganisation that is required for catalysis. Hydrolysis occurs via nucleophilic attack on the terminal phosphate. Finally, release of the metal ion allows the ‘open’ conformation to be restored for subsequent catalysis to occur.
Advisors/Committee Members: Arcus, Vickery L (advisor), Roberts, Nick (advisor).
Subjects/Keywords: NTPDase;
ATP
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APA (6th Edition):
Cumming, M. H. (2012). Structural and enzymatic characterisation of nucleoside triphosphate diphosphohydrolases from Trifolium repens and Dolichos biflorus
. (Doctoral Dissertation). University of Waikato. Retrieved from http://hdl.handle.net/10289/6331
Chicago Manual of Style (16th Edition):
Cumming, Mathew Hoani. “Structural and enzymatic characterisation of nucleoside triphosphate diphosphohydrolases from Trifolium repens and Dolichos biflorus
.” 2012. Doctoral Dissertation, University of Waikato. Accessed January 22, 2020.
http://hdl.handle.net/10289/6331.
MLA Handbook (7th Edition):
Cumming, Mathew Hoani. “Structural and enzymatic characterisation of nucleoside triphosphate diphosphohydrolases from Trifolium repens and Dolichos biflorus
.” 2012. Web. 22 Jan 2020.
Vancouver:
Cumming MH. Structural and enzymatic characterisation of nucleoside triphosphate diphosphohydrolases from Trifolium repens and Dolichos biflorus
. [Internet] [Doctoral dissertation]. University of Waikato; 2012. [cited 2020 Jan 22].
Available from: http://hdl.handle.net/10289/6331.
Council of Science Editors:
Cumming MH. Structural and enzymatic characterisation of nucleoside triphosphate diphosphohydrolases from Trifolium repens and Dolichos biflorus
. [Doctoral Dissertation]. University of Waikato; 2012. Available from: http://hdl.handle.net/10289/6331
University of Cambridge
2.
Douglas, Corsten Perrie Louise Claire.
Studies of the assembly pathway of human ATP synthase.
Degree: PhD, 2017, University of Cambridge
URL: https://www.repository.cam.ac.uk/handle/1810/267744
;
https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.725532
► Human mitochondrial ATP synthase is an enzyme containing 18 unlike subunits located in the inner mitochondrial membrane (IMM), where the catalytic F1 domain extends into…
(more)
▼ Human mitochondrial ATP synthase is an enzyme containing 18 unlike subunits located in the inner mitochondrial membrane (IMM), where the catalytic F1 domain extends into the mitochondrial matrix and the FO domain, which contains the c8-ring rotor, the a-subunit and the supernumerary subunits, is anchored in the IMM. All the subunits, apart from the a- and A6L-subunits, are encoded in the nucleus and require transport into the mitochondria before being assembled. The a- and A6L-subunits are encoded on the mitochondrial genome. The respiratory complexes generate the proton motive force (PMF), which ATP synthase uses to generate ATP from ADP and Pi. Rotation of the α- and β-subunits with the central stalk γ-, δ- and ε-subunits is prevented by coupling the F1 domain to the FO domain via the peripheral stalk (the OSCP-, F6-, d- and b-subunits). ATP hydrolysis is prevented by the natural inhibitor of the enzyme, IF1, binding to the F1 domain. In addition to the aand, b-subunits, the FO domain contains the c8-ring and six supernumerary subunits not involved in the catalytic activity of ATP synthase. The roles of five of these subunits in the assembly of ATP synthase, the e-, f-, g-, DAPIT- and 6.8 kDa proteolipid-subunits, were investigated by suppressing or disrupting their expression individually. The e-subunit is the first of the supernumerary subunits to assemble, then the g-subunit followed by the f-, 6.8 kDa proteolipid- and DAPIT-subunits. All five supernumerary subunits investigated were required to facilitate the dimerisation and oligomerisation of ATP synthase. The e-, f- and g-subunits were found to be important for maintaining mitochondrial respiratory capacity.
Subjects/Keywords: 572; ATP synthase; ATP synthase assembly; ATP synthase oligomerisation; ATP synthase dimerisation; ATP synthase subunit composition; ATP synthase supernumerary subunits
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APA (6th Edition):
Douglas, C. P. L. C. (2017). Studies of the assembly pathway of human ATP synthase. (Doctoral Dissertation). University of Cambridge. Retrieved from https://www.repository.cam.ac.uk/handle/1810/267744 ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.725532
Chicago Manual of Style (16th Edition):
Douglas, Corsten Perrie Louise Claire. “Studies of the assembly pathway of human ATP synthase.” 2017. Doctoral Dissertation, University of Cambridge. Accessed January 22, 2020.
https://www.repository.cam.ac.uk/handle/1810/267744 ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.725532.
MLA Handbook (7th Edition):
Douglas, Corsten Perrie Louise Claire. “Studies of the assembly pathway of human ATP synthase.” 2017. Web. 22 Jan 2020.
Vancouver:
Douglas CPLC. Studies of the assembly pathway of human ATP synthase. [Internet] [Doctoral dissertation]. University of Cambridge; 2017. [cited 2020 Jan 22].
Available from: https://www.repository.cam.ac.uk/handle/1810/267744 ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.725532.
Council of Science Editors:
Douglas CPLC. Studies of the assembly pathway of human ATP synthase. [Doctoral Dissertation]. University of Cambridge; 2017. Available from: https://www.repository.cam.ac.uk/handle/1810/267744 ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.725532
University of Cambridge
3.
Douglas, Corsten Perrie Louise Claire.
Studies of the assembly pathway of human ATP synthase.
Degree: PhD, 2017, University of Cambridge
URL: https://www.repository.cam.ac.uk/handle/1810/267744
► Human mitochondrial ATP synthase is an enzyme containing 18 unlike subunits located in the inner mitochondrial membrane (IMM), where the catalytic F1 domain extends into…
(more)
▼ Human mitochondrial ATP synthase is an enzyme containing 18 unlike subunits located in
the inner mitochondrial membrane (IMM), where the catalytic F1 domain extends into the
mitochondrial matrix and the FO domain, which contains the c8-ring rotor, the a-subunit
and the supernumerary subunits, is anchored in the IMM. All the subunits, apart from the
a- and A6L-subunits, are encoded in the nucleus and require transport into the mitochondria
before being assembled. The a- and A6L-subunits are encoded on the mitochondrial
genome. The respiratory complexes generate the proton motive force (PMF), which ATP
synthase uses to generate ATP from ADP and Pi. Rotation of the α- and β-subunits with the
central stalk γ-, δ- and ε-subunits is prevented by coupling the F1 domain to the FO domain
via the peripheral stalk (the OSCP-, F6-, d- and b-subunits). ATP hydrolysis is prevented
by the natural inhibitor of the enzyme, IF1, binding to the F1 domain. In addition to the aand,
b-subunits, the FO domain contains the c8-ring and six supernumerary subunits not
involved in the catalytic activity of ATP synthase. The roles of five of these subunits in the
assembly of ATP synthase, the e-, f-, g-, DAPIT- and 6.8 kDa proteolipid-subunits, were
investigated by suppressing or disrupting their expression individually. The e-subunit is the
first of the supernumerary subunits to assemble, then the g-subunit followed by the f-, 6.8
kDa proteolipid- and DAPIT-subunits. All five supernumerary subunits investigated were
required to facilitate the dimerisation and oligomerisation of ATP synthase. The e-, f- and
g-subunits were found to be important for maintaining mitochondrial respiratory capacity.
Subjects/Keywords: ATP synthase; ATP synthase assembly; ATP synthase oligomerisation; ATP synthase dimerisation; ATP synthase subunit composition; ATP synthase supernumerary subunits
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APA ·
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MLA ·
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Manager
APA (6th Edition):
Douglas, C. P. L. C. (2017). Studies of the assembly pathway of human ATP synthase. (Doctoral Dissertation). University of Cambridge. Retrieved from https://www.repository.cam.ac.uk/handle/1810/267744
Chicago Manual of Style (16th Edition):
Douglas, Corsten Perrie Louise Claire. “Studies of the assembly pathway of human ATP synthase.” 2017. Doctoral Dissertation, University of Cambridge. Accessed January 22, 2020.
https://www.repository.cam.ac.uk/handle/1810/267744.
MLA Handbook (7th Edition):
Douglas, Corsten Perrie Louise Claire. “Studies of the assembly pathway of human ATP synthase.” 2017. Web. 22 Jan 2020.
Vancouver:
Douglas CPLC. Studies of the assembly pathway of human ATP synthase. [Internet] [Doctoral dissertation]. University of Cambridge; 2017. [cited 2020 Jan 22].
Available from: https://www.repository.cam.ac.uk/handle/1810/267744.
Council of Science Editors:
Douglas CPLC. Studies of the assembly pathway of human ATP synthase. [Doctoral Dissertation]. University of Cambridge; 2017. Available from: https://www.repository.cam.ac.uk/handle/1810/267744
Arizona State University
4.
Yang, Jay-How.
Isolation and Functional Studies of The F-type ATP Synthase
from Spinach Chloroplasts and Heliobacterium modesticaldum.
Degree: Biochemistry, 2015, Arizona State University
URL: http://repository.asu.edu/items/29992
► Adenosine triphosphate (ATP) is the universal chemical energy currency in most living cells, used to power many cellular reactions and generated by an enzyme supercomplex…
(more)
▼ Adenosine triphosphate (ATP) is the universal chemical
energy currency in most living cells, used to power many cellular
reactions and generated by an enzyme supercomplex known as the ATP
synthase, consisting of a hydrophilic F1 subcomplex and a
membrane-bound FO subcomplex. Driven by the electrochemical
gradient generated by the respiratory or photosynthetic electron
transport chain, the rotation of the FO domain drives movements of
the central stalk in response to conformational changes in the F1
domain, in which the physical energy is converted into chemical
energy through the condensation of ADP and Pi to ATP. The exact
mechanism how ATP synthesis is coupled to proton translocation is
not known as no structure of the intact ATP-synthase nor the intact
FO subcomplex has been determined to date. Structural information
may shed light on these mechanisms and aid in understanding how
structural changed relate to its coupling to ATP synthesis. The
work in this thesis has successful established a defined
large-scale CF1FO isolation procedure resulting in high purity and
high yield of this complex from spinach thylakoid membranes by
incorporating a unique combination of biochemical methods will form
the basis for the subsequent structural determination of this
complex. Isolation began from the isolation of intact chloroplasts
and the separation of intact thylakoid membranes. Both native and
denaturing electrophoresis analyses clearly demonstrated that the
purified CF1FO retains its quaternary structure consisting of the
CF1 and CFO subcomplexes and nine subunits (five F1 subunits: α, β,
γ, δ and ε, and four FO subunits: a, b, b' and c). Moreover, both
ATP synthesis and hydrolysis activities were successfully detected
using protein reconstitution in combination with acid-base
incubation and in-gel ATPase assays, respectively. Furthermore, the
ATP-synthase of H. modesticaldum, an anaerobic photosynthetic
bacterium, was also isolated and characterized at the biochemical
level. These biochemical characterizations directly influenced
recent studies on the high-resolution structure determination of
intact CF1FO using electron crystallography on two-dimensional
crystals. The availability of the functionally intact CF1FO
purified at a large scale will lead to studies that investigate the
possible crystallization conditions to ultimately determine its
three-dimensional structure at atomic resolution.
Subjects/Keywords: Biochemistry; ATP synthase
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APA ·
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APA (6th Edition):
Yang, J. (2015). Isolation and Functional Studies of The F-type ATP Synthase
from Spinach Chloroplasts and Heliobacterium modesticaldum. (Doctoral Dissertation). Arizona State University. Retrieved from http://repository.asu.edu/items/29992
Chicago Manual of Style (16th Edition):
Yang, Jay-How. “Isolation and Functional Studies of The F-type ATP Synthase
from Spinach Chloroplasts and Heliobacterium modesticaldum.” 2015. Doctoral Dissertation, Arizona State University. Accessed January 22, 2020.
http://repository.asu.edu/items/29992.
MLA Handbook (7th Edition):
Yang, Jay-How. “Isolation and Functional Studies of The F-type ATP Synthase
from Spinach Chloroplasts and Heliobacterium modesticaldum.” 2015. Web. 22 Jan 2020.
Vancouver:
Yang J. Isolation and Functional Studies of The F-type ATP Synthase
from Spinach Chloroplasts and Heliobacterium modesticaldum. [Internet] [Doctoral dissertation]. Arizona State University; 2015. [cited 2020 Jan 22].
Available from: http://repository.asu.edu/items/29992.
Council of Science Editors:
Yang J. Isolation and Functional Studies of The F-type ATP Synthase
from Spinach Chloroplasts and Heliobacterium modesticaldum. [Doctoral Dissertation]. Arizona State University; 2015. Available from: http://repository.asu.edu/items/29992
University of Alberta
5.
Lai, Aaron.
Determining factors in the differential activation of
microglia.
Degree: PhD, Centre for Neuroscience, 2010, University of Alberta
URL: https://era.library.ualberta.ca/files/tb09j6275
► Microglia, the resident immune cells of the central nervous system (CNS), become activated in response to danger signals given out by other cells when homeostasis…
(more)
▼ Microglia, the resident immune cells of the central
nervous system (CNS), become activated in response to danger
signals given out by other cells when homeostasis has been
disturbed. Microglial activation is a multifaceted phenomenon that
includes numerous distinct phenotypes. The type of activation often
influences the survival of surrounding CNS tissue, and thus gaining
a better understanding of how microglial activation is regulated
has important therapeutic implications. Currently, it is known that
the phenotype of activated microglia depends on both the type of
CNS insult and the specific activating agent. The aim of this
thesis was to investigate the potential involvement of other
determining factors. Extrinsic regulators of microglial activation,
including the severity of CNS insult and the stimulation strength
of activating agents, were examined. Intrinsic differences among
different microglial populations, namely differences in region of
origin and age of origin, were also investigated. To study
microglial behavior without interference from other cells, rat
primary cultures were used as the system of study. With regard to
extrinsic factors, it was found that different severities of
hypoxic neuronal injury induced distinct microglial phenotypes.
Among the activating agents released by injured neurons, adenosine
5’-triphosphate (ATP) was studied in isolation and was found to
induce trophic and toxic effectors in microglia depending on the
strength of ATP stimulation. In regards to intrinsic factors, it
was found that microglia derived from different regions of the
brain had distinct responses to activators, with cortical and
hippocampal microglia generating more toxic responses than
brainstem, striatal, and thalamic microglia. Microglia derived from
various ages of origin also responded differentially to activators,
with neonatal and aged microglia being more reactive than microglia
derived from other age groups. Together, the results here present
several novel concepts, that the phenotype of activated microglia
are dependent not only on the type of activating stimulus, but the
strength of that stimulus, and that in addition to stimuli from
other cells, the regional and age differences among microglia
themselves are also crucial in determining their activation
phenotype.
Subjects/Keywords: ATP; hypoxia; neuroinflammation; neuroscience; microglia
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APA (6th Edition):
Lai, A. (2010). Determining factors in the differential activation of
microglia. (Doctoral Dissertation). University of Alberta. Retrieved from https://era.library.ualberta.ca/files/tb09j6275
Chicago Manual of Style (16th Edition):
Lai, Aaron. “Determining factors in the differential activation of
microglia.” 2010. Doctoral Dissertation, University of Alberta. Accessed January 22, 2020.
https://era.library.ualberta.ca/files/tb09j6275.
MLA Handbook (7th Edition):
Lai, Aaron. “Determining factors in the differential activation of
microglia.” 2010. Web. 22 Jan 2020.
Vancouver:
Lai A. Determining factors in the differential activation of
microglia. [Internet] [Doctoral dissertation]. University of Alberta; 2010. [cited 2020 Jan 22].
Available from: https://era.library.ualberta.ca/files/tb09j6275.
Council of Science Editors:
Lai A. Determining factors in the differential activation of
microglia. [Doctoral Dissertation]. University of Alberta; 2010. Available from: https://era.library.ualberta.ca/files/tb09j6275
University of Alberta
6.
Zwicker, Jennifer D.
ATP and central respiratory control: a three-part signaling
system.
Degree: PhD, Department of Physiology, 2012, University of Alberta
URL: https://era.library.ualberta.ca/files/ws859g94q
► ATP actions on central inspiratory networks are determined by a three-part signaling system comprising: i) excitatory actions of ATP at P2 receptors (Rs) ii) ectonucleotidases…
(more)
▼ ATP actions on central inspiratory networks are
determined by a three-part signaling system comprising: i)
excitatory actions of ATP at P2 receptors (Rs) ii)
ectonucleotidases that degrade ATP into adenosine (ADO), and iii)
inhibitory actions of ADO at P1Rs. During hypoxia, an initial
increase in ventilation is followed by a secondary depression that
is life threatening in premature infants. The release of ATP in
respiratory networks, including the preBötzinger Complex (preBötC,
inspiratory rhythm generation site) attenuates this secondary
ventilatory depression. However, subsequent degradation of ATP to
ADO may exacerbate the depression. The objective of my thesis
research is to explore the significance of this three-part
signaling system for preBötC networks in rodents using rhythmically
active medullary slices from rats and mice, primary cultures of
preBötC glia, and anesthetized adult rats. In neonatal rats in
vitro, injection of ATP into the preBötC evokes an increase in
inspiratory breathing frequency via a P2Y1R mechanism that involves
both neurons and glia. Analysis of cultured preBötC glia suggests
that P2Y1R stimulation evokes an increase in Ca2+ and release of
glutamate, which excites inspiratory neurons. In contrast,
injection of ATP into the preBötC of rhythmic slices from neonatal
mice evokes a P2Y1R-mediated frequency increase if A1 ADORs are
blocked. In contrast to rats, neonatal mice are sensitive to ADO
inhibition of preBötC frequency and have higher expression of the
ectonucleotidase TNAP (an enzyme that degrades ATP to ADO). A
delicate balance between P2R actions and P1R actions modulates the
preBötC network of neonatal rodents. Purinergic signaling also
influences the activity of mature preBötC networks in adult rats.
Injection of a P2Y1R agonist into the preBötC evoked a 40% increase
in respiratory frequency while ADO injection had no effect. As
predicted based on the proposed role of ATP in attenuating the
secondary hypoxic ventilatory depression, the increase of
endogenous ectonucleotidase activity in the preBötC (via injection
of a lentivirus controlling expression of the enzyme TMPAP)
produced a greater secondary hypoxic ventilatory depression
compared to control. This work lays the foundation for future
research examining the importance of glia and purinergic modulation
within the rhythmogenic inspiratory network.
Subjects/Keywords: ATP; PreBotzinger Complex; Glia; Adenosine
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APA ·
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Manager
APA (6th Edition):
Zwicker, J. D. (2012). ATP and central respiratory control: a three-part signaling
system. (Doctoral Dissertation). University of Alberta. Retrieved from https://era.library.ualberta.ca/files/ws859g94q
Chicago Manual of Style (16th Edition):
Zwicker, Jennifer D. “ATP and central respiratory control: a three-part signaling
system.” 2012. Doctoral Dissertation, University of Alberta. Accessed January 22, 2020.
https://era.library.ualberta.ca/files/ws859g94q.
MLA Handbook (7th Edition):
Zwicker, Jennifer D. “ATP and central respiratory control: a three-part signaling
system.” 2012. Web. 22 Jan 2020.
Vancouver:
Zwicker JD. ATP and central respiratory control: a three-part signaling
system. [Internet] [Doctoral dissertation]. University of Alberta; 2012. [cited 2020 Jan 22].
Available from: https://era.library.ualberta.ca/files/ws859g94q.
Council of Science Editors:
Zwicker JD. ATP and central respiratory control: a three-part signaling
system. [Doctoral Dissertation]. University of Alberta; 2012. Available from: https://era.library.ualberta.ca/files/ws859g94q
University of Minnesota
7.
Robinson, Jerid.
Activation and Inhibition Mechanisms of Membrane Guanylyl Cyclases.
Degree: PhD, Pharmacology, 2013, University of Minnesota
URL: http://hdl.handle.net/11299/173946
► Guanylyl cyclase (GC)-A and GC-B are homologous enzymes that catalyze the formation of cyclic guanosine monophosphate and pyrophosphate from GTP. GC-A is activated by atrial…
(more)
▼ Guanylyl cyclase (GC)-A and GC-B are homologous enzymes that catalyze the formation of cyclic guanosine monophosphate and pyrophosphate from GTP. GC-A is activated by atrial natriuretic peptide and B-type natriuretic peptide and regulates the cardiovascular system. GC-B is activated by C-type natriuretic peptide and regulates the skeletal and female reproductive systems. Activation of GC-A and GC-B was hypothesized to occur through two steps, binding of natriuretic peptide and subsequent binding of ATP to the kinase homology domain. However, our group reported that ATP binding does not increase maximal velocity but reduces the Michaelis constant. My work revealed an allosteric ATP binding site in the catalytic domain. Mutation and structure/function studies indicated that the allosteric and catalytic sites are different and that GC-A and GC-B are asymmetric homodimers, not symmetric homodimers as had been previously suggested. Interestingly, a constitutively active mutant of GC-B mimicked an ATP bound state. ATP inhibits soluble guanylyl cyclases (sGC), and I demonstrated that physiological concentrations of ATP inhibit GC-A and GC-B. I went on to determine that the mechanism of inhibition was through binding the pyrophosphate-product site. Together, these data revealed how low and high concentrations of ATP activate and inhibit GC-A and GC-B, respectively.
Subjects/Keywords: ANP; ATP; Guanylyl Cyclase
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APA (6th Edition):
Robinson, J. (2013). Activation and Inhibition Mechanisms of Membrane Guanylyl Cyclases. (Doctoral Dissertation). University of Minnesota. Retrieved from http://hdl.handle.net/11299/173946
Chicago Manual of Style (16th Edition):
Robinson, Jerid. “Activation and Inhibition Mechanisms of Membrane Guanylyl Cyclases.” 2013. Doctoral Dissertation, University of Minnesota. Accessed January 22, 2020.
http://hdl.handle.net/11299/173946.
MLA Handbook (7th Edition):
Robinson, Jerid. “Activation and Inhibition Mechanisms of Membrane Guanylyl Cyclases.” 2013. Web. 22 Jan 2020.
Vancouver:
Robinson J. Activation and Inhibition Mechanisms of Membrane Guanylyl Cyclases. [Internet] [Doctoral dissertation]. University of Minnesota; 2013. [cited 2020 Jan 22].
Available from: http://hdl.handle.net/11299/173946.
Council of Science Editors:
Robinson J. Activation and Inhibition Mechanisms of Membrane Guanylyl Cyclases. [Doctoral Dissertation]. University of Minnesota; 2013. Available from: http://hdl.handle.net/11299/173946
Addis Ababa University
8.
Asmamaw, Asrat.
DYNAMICS OF MOTOR PROTEINS= A MONTE CARLO SIMULATION
.
Degree: 2012, Addis Ababa University
URL: http://etd.aau.edu.et/dspace/handle/123456789/1190
► Motor proteins are mechanochemical enzymes that convert energy released by adenosine triphosphate (ATP) hydrolysis into either linear or rotary movement. Motor proteins: kinesin, myosin and…
(more)
▼ Motor proteins are mechanochemical enzymes that convert energy released by adenosine
triphosphate (
ATP) hydrolysis into either linear or rotary movement. Motor proteins:
kinesin, myosin and dynein, that perform active movements along cytoskeletal laments
drive the long=range transport of vesicles, organelles, and other types of cargo in biological
cells. In this Monte Carlo simulation study, based on the recent experimental results and
existing theoretical models, a lattice model to study the dynamics of non=interacting
motor proteins each transporting a cargo and a new bead=spring model to study the
collective dynamics of interacting motor proteins transporting cooperatively a common
cargo are proposed and studied.
ix
Advisors/Committee Members: Dr. Tatek Yergou (advisor).
Subjects/Keywords: MOTOR PROTEINS;
adenosine triphosphate (ATP)
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APA ·
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to Zotero / EndNote / Reference
Manager
APA (6th Edition):
Asmamaw, A. (2012). DYNAMICS OF MOTOR PROTEINS= A MONTE CARLO SIMULATION
. (Thesis). Addis Ababa University. Retrieved from http://etd.aau.edu.et/dspace/handle/123456789/1190
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):
Asmamaw, Asrat. “DYNAMICS OF MOTOR PROTEINS= A MONTE CARLO SIMULATION
.” 2012. Thesis, Addis Ababa University. Accessed January 22, 2020.
http://etd.aau.edu.et/dspace/handle/123456789/1190.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Asmamaw, Asrat. “DYNAMICS OF MOTOR PROTEINS= A MONTE CARLO SIMULATION
.” 2012. Web. 22 Jan 2020.
Vancouver:
Asmamaw A. DYNAMICS OF MOTOR PROTEINS= A MONTE CARLO SIMULATION
. [Internet] [Thesis]. Addis Ababa University; 2012. [cited 2020 Jan 22].
Available from: http://etd.aau.edu.et/dspace/handle/123456789/1190.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Asmamaw A. DYNAMICS OF MOTOR PROTEINS= A MONTE CARLO SIMULATION
. [Thesis]. Addis Ababa University; 2012. Available from: http://etd.aau.edu.et/dspace/handle/123456789/1190
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Ruhr Universität Bochum
9.
Veitinger, Thomas.
Physiologische Untersuchungen von Signalwegen in reifen
und unreifen Keimzellen männlicher Säugetiere.
Degree: 2009, Ruhr Universität Bochum
URL: http://nbn-resolving.de/urn/resolver.pl?urn=urn:nbn:de:hbz:294-30273
► In der vorliegenden Arbeit wurden physiologische Reaktionen von menschlichen Spermien im Zusammenhang mit Chemotaxis und Motilität untersucht. Dabei wurden Spermien verschiedenen Duftstoffen ausgesetzt und mit…
(more)
▼ In der vorliegenden Arbeit wurden physiologische
Reaktionen von menschlichen Spermien im Zusammenhang mit Chemotaxis
und Motilität untersucht. Dabei wurden Spermien verschiedenen
Duftstoffen ausgesetzt und mit Hilfe bildgebender Verfahren die
intrazelluläre Kalziumdynamik gemessen. Zusätzlich wurde die
Akkumulation von Molekülen wie z.B.
ATP mittels ELISA
quantifiziert. Weiterhin wurden in einem Kokulturansatz die
ATP-
und Hormonsignalgebung muriner Spermatogonien untersucht. Hierfür
wurden kultivierte murine Spermatogonien verschiedenen Hormonen und
ATP-Konzentrationen ausgesetzt und das Antwortverhalten mit Hilfe
bildgebender Verfahren aufgezeichnet.
Advisors/Committee Members: Biologie.
Subjects/Keywords: Spermium; Calcium; ATP; Spermatogenese;
Chemotaxis
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APA (6th Edition):
Veitinger, T. (2009). Physiologische Untersuchungen von Signalwegen in reifen
und unreifen Keimzellen männlicher Säugetiere. (Thesis). Ruhr Universität Bochum. Retrieved from http://nbn-resolving.de/urn/resolver.pl?urn=urn:nbn:de:hbz:294-30273
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):
Veitinger, Thomas. “Physiologische Untersuchungen von Signalwegen in reifen
und unreifen Keimzellen männlicher Säugetiere.” 2009. Thesis, Ruhr Universität Bochum. Accessed January 22, 2020.
http://nbn-resolving.de/urn/resolver.pl?urn=urn:nbn:de:hbz:294-30273.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Veitinger, Thomas. “Physiologische Untersuchungen von Signalwegen in reifen
und unreifen Keimzellen männlicher Säugetiere.” 2009. Web. 22 Jan 2020.
Vancouver:
Veitinger T. Physiologische Untersuchungen von Signalwegen in reifen
und unreifen Keimzellen männlicher Säugetiere. [Internet] [Thesis]. Ruhr Universität Bochum; 2009. [cited 2020 Jan 22].
Available from: http://nbn-resolving.de/urn/resolver.pl?urn=urn:nbn:de:hbz:294-30273.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Veitinger T. Physiologische Untersuchungen von Signalwegen in reifen
und unreifen Keimzellen männlicher Säugetiere. [Thesis]. Ruhr Universität Bochum; 2009. Available from: http://nbn-resolving.de/urn/resolver.pl?urn=urn:nbn:de:hbz:294-30273
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Case Western Reserve University
10.
Ward, Jessica.
Deciphering the Proteolytic Mechanism of the ATP-Dependent
Protease Lon Using Fluorescent Peptides.
Degree: PhD, Chemistry, 2008, Case Western Reserve University
URL: http://rave.ohiolink.edu/etdc/view?acc_num=case1194311050
► Lon is an ATP-dependent serine protease that degrades damaged, misfolded and certain regulatory proteins in the cell. The enzyme exists as a homo-oligomer with one…
(more)
▼ Lon is an
ATP-dependent serine protease that degrades
damaged, misfolded and certain regulatory proteins in the cell. The
enzyme exists as a homo-oligomer with one ATPase domain and one
protease domain on each subunit. Using our fluorescent model
peptide substrate, ¿N89-98, as a tool, we have employed
steady-state and pre-steady-state kinetic techniques to evaluate
the proteolytic mechanism of Lon protease. Steady-state kinetic
analysis of the peptidase activity of Lon with nucleotide
triphosphates reveals that while nucleotide binding alone is enough
to support peptide hydrolysis, the contribution of nucleotide
hydrolysis towards the proteolytic activity is significant.
Pre-steady-state experiments reveal a lag phase in the peptidase
reaction and that
ATP hydrolysis occurs before the peptide cleavage
event. We propose that the slow step constitues a peptide
translocation event that contains two binding events in which the
peptide is delivered from the initial peptide binding site to the
protease active site. I was able to measure this slow step using
proteolytic inactive Lon mutants and a dansylated version of the
¿N89-98 peptide as well. Taken together, I propose a mechanism for
the first round of peptide hydrolysis. Following initial nucleotide
and peptide binding, a nucleotide dependent conformational change
occurs. Next, upon
ATP hydrolysis, the proteolytic site is
activated and peptide is delivered to the protease domain where
peptide cleavage occurs. In addition to characterizing the kinetic
mechanism of Lon protease, I have also utilized multiple peptide
substrates to demonstrate the importance of the P3, P1 and P3’
positions in determining substrate specificity and provide evidence
that the C-terminus of the ¿N protein is important for recognition
by Lon.
Advisors/Committee Members: Zagorski, Michael (Committee Chair).
Subjects/Keywords: LON; ATP; PEPTIDES; S679W; ATPase
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APA (6th Edition):
Ward, J. (2008). Deciphering the Proteolytic Mechanism of the ATP-Dependent
Protease Lon Using Fluorescent Peptides. (Doctoral Dissertation). Case Western Reserve University. Retrieved from http://rave.ohiolink.edu/etdc/view?acc_num=case1194311050
Chicago Manual of Style (16th Edition):
Ward, Jessica. “Deciphering the Proteolytic Mechanism of the ATP-Dependent
Protease Lon Using Fluorescent Peptides.” 2008. Doctoral Dissertation, Case Western Reserve University. Accessed January 22, 2020.
http://rave.ohiolink.edu/etdc/view?acc_num=case1194311050.
MLA Handbook (7th Edition):
Ward, Jessica. “Deciphering the Proteolytic Mechanism of the ATP-Dependent
Protease Lon Using Fluorescent Peptides.” 2008. Web. 22 Jan 2020.
Vancouver:
Ward J. Deciphering the Proteolytic Mechanism of the ATP-Dependent
Protease Lon Using Fluorescent Peptides. [Internet] [Doctoral dissertation]. Case Western Reserve University; 2008. [cited 2020 Jan 22].
Available from: http://rave.ohiolink.edu/etdc/view?acc_num=case1194311050.
Council of Science Editors:
Ward J. Deciphering the Proteolytic Mechanism of the ATP-Dependent
Protease Lon Using Fluorescent Peptides. [Doctoral Dissertation]. Case Western Reserve University; 2008. Available from: http://rave.ohiolink.edu/etdc/view?acc_num=case1194311050
Louisiana State University
11.
Hendrix, Amanda Yoesting.
Base-caged Adenosine Triphosphate as a Model System for Photoactivatable Small Interfering RNA.
Degree: MSBAE, Engineering, 2012, Louisiana State University
URL: etd-06062012-164102
;
https://digitalcommons.lsu.edu/gradschool_theses/527
► Photocaged adenosine triphosphate (ATP) is one of the earliest examples of exerting spatial-temporal control over the activity of a substrate. The activity of ATP is…
(more)
▼ Photocaged adenosine triphosphate (ATP) is one of the earliest examples of exerting spatial-temporal control over the activity of a substrate. The activity of ATP is blocked until near-ultraviolet light exposure photocleaves the cage moiety. Caged ATP has been used for a myriad of applications including kinetic studies of ATP-dependent enzymes. Traditional caging of ATP occurs at the gamma-phosphate, which has been found to competitively inhibit several enzymatic systems. It was hypothesized that blocking access to the adenosine N6 position via cage molecule would prevent the initial enzyme-substrate binding event from occurring prior to photolysis, effectively minimizing competitive inhibition. Utilizing a convertible nucleoside analog of ATP, this work synthesized, purified, and characterized a form of caged ATP which, by attaching the cage molecule to the nucleobase, did not inhibit the enzymatic activity of luciferase in vitro. Characterization was accomplished via UV/Vis spectroscopy, high performance liquid chromatography (HPLC), nuclear magnetic resonance (NMR), and mass spectrometry (MS). Base-caged ATP was evaluated in a firefly luciferase enzymatic assay to determine the degree of bioactivity in the caged and photoactivated states and compared to the results of native (uncaged) ATP and gamma-NPE-caged ATP. Photolysis was conducted via 308 nm light from a transilluminator. Base-caged ATP did not inhibit the enzymatic system and the convertible nucleoside synthesis approach offers significant advantages over other caging techniques.
Subjects/Keywords: competitive inhibition; photoactivation; ATP; caged
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APA (6th Edition):
Hendrix, A. Y. (2012). Base-caged Adenosine Triphosphate as a Model System for Photoactivatable Small Interfering RNA. (Masters Thesis). Louisiana State University. Retrieved from etd-06062012-164102 ; https://digitalcommons.lsu.edu/gradschool_theses/527
Chicago Manual of Style (16th Edition):
Hendrix, Amanda Yoesting. “Base-caged Adenosine Triphosphate as a Model System for Photoactivatable Small Interfering RNA.” 2012. Masters Thesis, Louisiana State University. Accessed January 22, 2020.
etd-06062012-164102 ; https://digitalcommons.lsu.edu/gradschool_theses/527.
MLA Handbook (7th Edition):
Hendrix, Amanda Yoesting. “Base-caged Adenosine Triphosphate as a Model System for Photoactivatable Small Interfering RNA.” 2012. Web. 22 Jan 2020.
Vancouver:
Hendrix AY. Base-caged Adenosine Triphosphate as a Model System for Photoactivatable Small Interfering RNA. [Internet] [Masters thesis]. Louisiana State University; 2012. [cited 2020 Jan 22].
Available from: etd-06062012-164102 ; https://digitalcommons.lsu.edu/gradschool_theses/527.
Council of Science Editors:
Hendrix AY. Base-caged Adenosine Triphosphate as a Model System for Photoactivatable Small Interfering RNA. [Masters Thesis]. Louisiana State University; 2012. Available from: etd-06062012-164102 ; https://digitalcommons.lsu.edu/gradschool_theses/527
12.
Achterberg, Peter.
Formation and breakdown of adenosine in the heart : investigations on myocardial purine metabolismen.
Degree: 1986, Erasmus University Medical Center
URL: http://hdl.handle.net/1765/39037
► textabstractAdenosine, a strong coronary vasodilator, is a breakdown product of the myocardial high-energy phosphate ATP. ATP serves as the direct energy source for contraction of…
(more)
▼ textabstractAdenosine, a strong coronary vasodilator, is a breakdown
product of the myocardial high-energy phosphate ATP. ATP serves as
the direct energy source for contraction of the heart. Chapter 1 of
this thesis gives a general introduction on contractility dependent
ATP-breakdown, the ATP-generating metabolism of the heart and the
mechanism which leads to adenosine formation. The applications of
purine biochemistry to cardiovascular research are summarized. The in vitro characteristics of other enzymes of purine
metabolism (AMP-deaminase and S-adenosylhomocysteine hydrolase) are
reported. Additional perfusion studies with specific inhibitors
suggest a significant contribution by these enzymes to myocardial
purine formation during normoxia (Appendix Papers II and IV) .
Several lines of evidence suggest that the purines released from
isolated rat hearts are in part degradation products of IMP or GMP
instead of AMP. This appears to be especially so during normoxia.
If hypoxanthine or inosine are added to the myocardial
perfusate, they are slowly incorporated into the ATP-pool of the
heart. The incorporation rate is increased after a previous
ischemic period and can be further stimulated by simultaneous
infusion of ribose.
Evidence is presented that increased adenosine formation is
directly related to increased formation of AMP (e.g., from ATP).
The old hypothesis that adenosine is constantly produced at a high
rate and nearly simultaneously reincorporated into AMP appears to
be unrealistic. The results suggest an even tighter coupling of
adenosine and purine release to the myocardial energy state than
had already been assumed. A minor disbalance between ATP-breakdown
and ATP-production will lead to increased adenosine and purine
release and will eventually lead to significant loss of ATP and
myocardial function. Some of the experiments point to ways of
breaking through this vicious circle by defining conditions for
enhanced myocardial ATP-repletion and possible restoration of
function during reperfusion
Subjects/Keywords: ATP; adenosine
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MLA ·
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CSE |
Export
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Manager
APA (6th Edition):
Achterberg, P. (1986). Formation and breakdown of adenosine in the heart : investigations on myocardial purine metabolismen. (Doctoral Dissertation). Erasmus University Medical Center. Retrieved from http://hdl.handle.net/1765/39037
Chicago Manual of Style (16th Edition):
Achterberg, Peter. “Formation and breakdown of adenosine in the heart : investigations on myocardial purine metabolismen.” 1986. Doctoral Dissertation, Erasmus University Medical Center. Accessed January 22, 2020.
http://hdl.handle.net/1765/39037.
MLA Handbook (7th Edition):
Achterberg, Peter. “Formation and breakdown of adenosine in the heart : investigations on myocardial purine metabolismen.” 1986. Web. 22 Jan 2020.
Vancouver:
Achterberg P. Formation and breakdown of adenosine in the heart : investigations on myocardial purine metabolismen. [Internet] [Doctoral dissertation]. Erasmus University Medical Center; 1986. [cited 2020 Jan 22].
Available from: http://hdl.handle.net/1765/39037.
Council of Science Editors:
Achterberg P. Formation and breakdown of adenosine in the heart : investigations on myocardial purine metabolismen. [Doctoral Dissertation]. Erasmus University Medical Center; 1986. Available from: http://hdl.handle.net/1765/39037
University of Debrecen
13.
Józsa, Réka Anna.
A [(η6-p-cym)Ru(H2O)3]2+ kölcsönhatása ATP-vel
.
Degree: DE – Természettudományi és Technológiai Kar – Kémiai Intézet, University of Debrecen
URL: http://hdl.handle.net/2437/226843
► Az ATP a DNS egyik alkotórésze, a négy nukleotid egyike, bizonyos esetekben az ATP DNS modellnek is tekinthető, így fiziológiás pH-n az esetlegesen kialakuló komplexek…
(more)
▼ Az
ATP a DNS egyik alkotórésze, a négy nukleotid egyike, bizonyos esetekben az
ATP DNS modellnek is tekinthető, így fiziológiás pH-n az esetlegesen kialakuló komplexek nagy stabilitásából arra következtethetünk, hogy a rákos sejtek DNS-ében is kialakulnak a komplexek, így megakadályozva a sejtek szaporodását, növekedését esetlegesen a sejtek pusztulásához is vezethetnek.
A ruténium, mint platinafém tulajdonságaiban hasonló a platinához, így a tudósok úgy vélik, hogy ruténium tartalmú készítmények vizsgálata érdekes új eredményeket hozhat a rák kezelésével kapcsolatos ismeretek bővítésében. Ennek okán vizsgáltam a [(η6-p-cym)Ru(H2O)3]2+-ion (továbbiakban Ru(II)ion) komplexképzését
ATP, azaz adenozin-trifoszfát ligandummal, pH-potenciometriás módszerrel, mely alapján a komplexek sztöchiometriája és a protonálódási állandókat határozzuk meg, majd ezt követően 1H-NMR és ESI-MS mérésekkel, melyek alapján a lehetséges oldatszerkezetekről kaphatunk képet.
Advisors/Committee Members: Buglyó, Péter (advisor).
Subjects/Keywords: Ruténium;
ATP
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APA (6th Edition):
Józsa, R. A. (n.d.). A [(η6-p-cym)Ru(H2O)3]2+ kölcsönhatása ATP-vel
. (Thesis). University of Debrecen. Retrieved from http://hdl.handle.net/2437/226843
Note: this citation may be lacking information needed for this citation format:
No year of publication.
Not specified: Masters Thesis or Doctoral Dissertation
Chicago Manual of Style (16th Edition):
Józsa, Réka Anna. “A [(η6-p-cym)Ru(H2O)3]2+ kölcsönhatása ATP-vel
.” Thesis, University of Debrecen. Accessed January 22, 2020.
http://hdl.handle.net/2437/226843.
Note: this citation may be lacking information needed for this citation format:
No year of publication.
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Józsa, Réka Anna. “A [(η6-p-cym)Ru(H2O)3]2+ kölcsönhatása ATP-vel
.” Web. 22 Jan 2020.
Note: this citation may be lacking information needed for this citation format:
No year of publication.
Vancouver:
Józsa RA. A [(η6-p-cym)Ru(H2O)3]2+ kölcsönhatása ATP-vel
. [Internet] [Thesis]. University of Debrecen; [cited 2020 Jan 22].
Available from: http://hdl.handle.net/2437/226843.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
No year of publication.
Council of Science Editors:
Józsa RA. A [(η6-p-cym)Ru(H2O)3]2+ kölcsönhatása ATP-vel
. [Thesis]. University of Debrecen; Available from: http://hdl.handle.net/2437/226843
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
No year of publication.
Wayne State University
14.
Dedigama Arachchige, Pavithra Maheshani.
Development Of Tools For Phosphosite-Specific Kinase Identification And Discovery Of Phosphatase Substrates.
Degree: PhD, Chemistry, 2017, Wayne State University
URL: https://digitalcommons.wayne.edu/oa_dissertations/1693
► Phosphorylation is a ubiquitous post translational modification implicated in many diseases, such as cancer. The phosphorylation status of cellular proteins is regulated by the…
(more)
▼ Phosphorylation is a ubiquitous post translational modification implicated in many diseases, such as cancer. The phosphorylation status of cellular proteins is regulated by the activity of kinases and phosphatases. The biological significance of many phosphorylation events remain unknown because the methods to determine which kinase or phosphatase is responsible for phosphorylation are limited. Previously, we established kinase-catalyzed labeling where kinases accept γ-modified
ATP analogs, such as
ATP-arylazide and
ATP-biotin, to label phosphoproteins. To study substrates of kinases and phosphatases, here we developed two new methods using kinase-catalyzed labeling. As one application, we developed K-CLASP (Kinase-catalyzed CrossLinking And Streptavdin Purification) to identify the in-cellulo kinase of a phosphorylated site on a protein. In this case, we used
ATP-arylazide to mediate crosslinking between a biotin tagged peptide carrying a phosphosite of interest and the respective kinase. Using Protein kinase A (PKA) and its known peptide substrate kemptide, we demonstrated that K-CLASP is capable of identifying PKA as the kinase responsible for kemptide phosphorylation in cell lysates. Then we used K-CLASP to identify the kinases that phosphorylate S178 of the Miz1 protein in a collaboration project. For phosphatase substrate identification, we developed K-BIPS (Kinase-catalyzed Biotinylation to Identify Phosphatase Substrates). In prior work, we observed that labeling of phosphoproteins by
ATP-biotin is reduced when phosphatases are inactive. The phosphatase dependency of biotinylation is due to the presence of already existing phosphorylation, which prevents
ATP-biotin labeling. Therefore, in K-BIPS,
ATP-biotin labeling is carried out after the inactivation of a particular phosphatase. The loss in biotinylation can then be analyzed to reveal substrates. To establish K-BIPS as a viable method, we carried out
ATP-biotin labeling in lysates treated with the general phosphatase inhibitor okadaic acid. Many known phosphatase substrates were observed validating our method. Then as further applications, we used K-BIPS to explore substrates of PP1-Gadd34 and PP1-MYPT1 phosphatase complexes. The results demonstrate that K-BIS-Phos is a feasible method for phosphatase substrate identification. In summary, we have developed two chemical tools based on kinase-catalyzed labeling that will enhance our understanding of phosphorylation events mediated by kinases and phosphatases.
Advisors/Committee Members: Mary Kay Pflum.
Subjects/Keywords: ATP analogs; Kinase; phosphatase; Biochemistry
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APA (6th Edition):
Dedigama Arachchige, P. M. (2017). Development Of Tools For Phosphosite-Specific Kinase Identification And Discovery Of Phosphatase Substrates. (Doctoral Dissertation). Wayne State University. Retrieved from https://digitalcommons.wayne.edu/oa_dissertations/1693
Chicago Manual of Style (16th Edition):
Dedigama Arachchige, Pavithra Maheshani. “Development Of Tools For Phosphosite-Specific Kinase Identification And Discovery Of Phosphatase Substrates.” 2017. Doctoral Dissertation, Wayne State University. Accessed January 22, 2020.
https://digitalcommons.wayne.edu/oa_dissertations/1693.
MLA Handbook (7th Edition):
Dedigama Arachchige, Pavithra Maheshani. “Development Of Tools For Phosphosite-Specific Kinase Identification And Discovery Of Phosphatase Substrates.” 2017. Web. 22 Jan 2020.
Vancouver:
Dedigama Arachchige PM. Development Of Tools For Phosphosite-Specific Kinase Identification And Discovery Of Phosphatase Substrates. [Internet] [Doctoral dissertation]. Wayne State University; 2017. [cited 2020 Jan 22].
Available from: https://digitalcommons.wayne.edu/oa_dissertations/1693.
Council of Science Editors:
Dedigama Arachchige PM. Development Of Tools For Phosphosite-Specific Kinase Identification And Discovery Of Phosphatase Substrates. [Doctoral Dissertation]. Wayne State University; 2017. Available from: https://digitalcommons.wayne.edu/oa_dissertations/1693
15.
Cline, Paul.
Female Airline Transport Pilots: The Role of Mentoring.
Degree: PhD, Aviation, 2017, University of North Dakota
URL: https://commons.und.edu/theses/344
► Women have been a part of aviation since its inception, yet they have been traditionally underrepresented in the ranks of commercial pilots. This study…
(more)
▼ Women have been a part of aviation since its inception, yet they have been traditionally underrepresented in the ranks of commercial pilots. This study explored what role mentoring played in the lives and careers of female Airline Transport Pilots (
ATP). Participants completed a modified version of the Mentor Role Instrument (MRI) developed by Ragins and McFarlin.
It was determined that there was no statistically significant difference between female
ATP who had been mentored and those who had not. Of the female
ATP who had been mentored, those who reported an informal mentoring relationship rated their relationship higher than those who reported a formal mentoring relationship when it came to career oriented assistance and advice. The results for mentoring factors related to psychosocial needs and activities are less certain, but the preponderance of evidence supports the assertion that those female
ATP who reported an informal mentoring relationship were more satisfied in these areas than their formal mentor counterparts.
Advisors/Committee Members: Dr. Kimberly Kenville.
Subjects/Keywords: Airlines; ATP; Female; Mentoring; Pilot
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APA (6th Edition):
Cline, P. (2017). Female Airline Transport Pilots: The Role of Mentoring. (Doctoral Dissertation). University of North Dakota. Retrieved from https://commons.und.edu/theses/344
Chicago Manual of Style (16th Edition):
Cline, Paul. “Female Airline Transport Pilots: The Role of Mentoring.” 2017. Doctoral Dissertation, University of North Dakota. Accessed January 22, 2020.
https://commons.und.edu/theses/344.
MLA Handbook (7th Edition):
Cline, Paul. “Female Airline Transport Pilots: The Role of Mentoring.” 2017. Web. 22 Jan 2020.
Vancouver:
Cline P. Female Airline Transport Pilots: The Role of Mentoring. [Internet] [Doctoral dissertation]. University of North Dakota; 2017. [cited 2020 Jan 22].
Available from: https://commons.und.edu/theses/344.
Council of Science Editors:
Cline P. Female Airline Transport Pilots: The Role of Mentoring. [Doctoral Dissertation]. University of North Dakota; 2017. Available from: https://commons.und.edu/theses/344
University of Illinois – Chicago
16.
Tchernookova, Boriana Krassimirova.
Characterization of Extracellular H+ Fluxes from Isolated Glia and Slices of the Vertebrate Retina.
Degree: 2017, University of Illinois – Chicago
URL: http://hdl.handle.net/10027/21812
► Active glia-neuron interactions are crucial for the proper development and function of the nervous system. Bidirectional communication between neurons and glial cells has been shown…
(more)
▼ Active glia-neuron interactions are crucial for the proper development and function of the nervous system. Bidirectional communication between neurons and glial cells has been shown to have modulatory effects on neuronal signaling. The exact mechanisms which glial cells employ to influence the behavior of neurons have not been fully understood. In the vertebrate retina, extracellular pH can act as a modulator of neuronal signaling by altering the activity of calcium channels, particularly on photoreceptor terminals. Experiments have demonstrated that when the pH of the extracellular medium is lowered to more acidic values, the calcium flux into photoreceptors is lessened. The exact cell type, which mediates these changes in extracellular pH, is still debatable with mounting evidence pointing to the role of horizontal cells in this process. This work presents evidence that retina Müller glial cells of tiger salamander are capable of mediating changes in extracellular proton concentration. Müller glia respond to extracellular
ATP with a robust extracellular acidification measured with ion-selective microelectrodes. The mechanism through which these cells change the extracellular acidity involves the activation of membrane
ATP receptors and the mobilization of intracellular signaling molecules with resulting intracellular calcium increases. The
ATP-induced sustained acidifications are observed from isolated Müller glia as well as in retinal slices. Interestingly, Müller cells of the retinae of many other vertebrates tested, including Macaques and human, also respond to extracellular
ATP through acidifications of the extracellular space, suggesting that this is a conserved and potentially physiologically important response. Müller cells respond to extracellular glutamate as well, but the changes in extracellular acidity are opposite of these evoked by
ATP: glutamate induces Müller cell-mediated extracellular transient alkalinization, which seems to be due to the activity of membrane glutamate transporters. In slices, the initial transient alkalinization induced by glutamate is followed by a pronounced, sustained acidification, whose mechanism is independent of the glutamate transporters’ activity. This data suggest Müller glia could potentially play a role in extracellular proton modulation of neuronal activity in the vertebrate retina
Advisors/Committee Members: Malchow, Robert P (advisor), Park, Thomas (committee member), Gong, Liang-Wei (committee member), Matthew Kreitzer (committee member), Murphy, Alvin D (chair).
Subjects/Keywords: retina; glia; ATP; pH
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Manager
APA (6th Edition):
Tchernookova, B. K. (2017). Characterization of Extracellular H+ Fluxes from Isolated Glia and Slices of the Vertebrate Retina. (Thesis). University of Illinois – Chicago. Retrieved from http://hdl.handle.net/10027/21812
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):
Tchernookova, Boriana Krassimirova. “Characterization of Extracellular H+ Fluxes from Isolated Glia and Slices of the Vertebrate Retina.” 2017. Thesis, University of Illinois – Chicago. Accessed January 22, 2020.
http://hdl.handle.net/10027/21812.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Tchernookova, Boriana Krassimirova. “Characterization of Extracellular H+ Fluxes from Isolated Glia and Slices of the Vertebrate Retina.” 2017. Web. 22 Jan 2020.
Vancouver:
Tchernookova BK. Characterization of Extracellular H+ Fluxes from Isolated Glia and Slices of the Vertebrate Retina. [Internet] [Thesis]. University of Illinois – Chicago; 2017. [cited 2020 Jan 22].
Available from: http://hdl.handle.net/10027/21812.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Tchernookova BK. Characterization of Extracellular H+ Fluxes from Isolated Glia and Slices of the Vertebrate Retina. [Thesis]. University of Illinois – Chicago; 2017. Available from: http://hdl.handle.net/10027/21812
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
17.
Julkunen, Henna.
Käyttöohje voilinjan CIP-pesuprosessista
.
Degree: 2011, Theseus
URL: http://www.theseus.fi/handle/10024/32275
► Opinnäytetyön tavoitteena oli tehdä yksityiskohtainen käyttöohje uuden rasvatehtaan voilinjan CIP- pesuprosessista ja sen toiminnasta sekä ohjauksesta. Käyttöohje tehtiin Valio Oy:n Seinäjoen rasvatehtaalle. Käyttöohjeen tarkoitus on…
(more)
▼ Opinnäytetyön tavoitteena oli tehdä yksityiskohtainen käyttöohje uuden rasvatehtaan voilinjan CIP- pesuprosessista ja sen toiminnasta sekä ohjauksesta. Käyttöohje tehtiin Valio Oy:n Seinäjoen rasvatehtaalle.
Käyttöohjeen tarkoitus on antaa selkeät toimintaohjeet turvalliseen pesutoimintoon sekä parhaan pesutuloksen saavuttamiseen. Käyttöohje opastaa rutiininomaisesti valvomon työntekijöitä ja yhtenäistää heidän toimintatapojaan. Käyttöohje antaa myös tukea itsenäiseen työskentelyyn valvomossa.
Ennen uuden tehtaan valmistumista pesuprosessin toiminnasta ja ohjauksesta vastasivat kokeneet valvomotyöntekijät. Uuden organisaatiomuutoksen myötä myös voinvalmistajat työskentelevät ajoittain valvomossa, joten käyttöohjeita eri toiminnoista pyydettiin tukemaan heidän valvomotyöskentelyään.
Käyttöohjeen lisäksi työssä perehdyttiin perusteellisemmin elintarvikkeita koskeviin hygieniavaatimuksiin sekä elintarviketeollisuuden kaupalliseen, moraaliseen ja lainmukaiseen velvoitteeseen. Työssä käsiteltiin myös käsitteet: steriili, bakteriologinen sekä kemiallinen puhtaus. Työssä käytiin läpi myös ATP- menetelmällä otettuja puhdistustuloksia voilinjalta, jonka pesuun käyttöohje on tehty.; The object for the thesis was to create a detailed operation manual for operators on CIP washing process in the new butter plant. The manual includes all functions and controls to operate CIP washing process that is used to clean manufacturing lines in the butter plant. The manual was made to Valio PLC, Seinäjoki.
The purpose of the manual is to provide clear instructions for a safe washing process and the best possible washing result. The manual is created to advise operators in the control room and to standardise their way of working. The manual is also made to provide support to operators in the control room.
Before the new plant was built, experienced control room operators were responsible for the control of the CIP washing process. Due to changes in the new organization, now butter makers are responsible for CIP washing process and a well detailed operation manual is needed.
In addition to the operation manual, the thesis studies also food hygiene and commercial, moral and legal obligation of food industry. Also the concepts sterile, bacteriological and chemical cleanliness are handled. The cleanliness tests from butter manufacturing lines taken with ATP- method are part of the thesis as well.
Subjects/Keywords: elintarvikehygienia;
pesu;
CIP;
ATP
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APA (6th Edition):
Julkunen, H. (2011). Käyttöohje voilinjan CIP-pesuprosessista
. (Thesis). Theseus. Retrieved from http://www.theseus.fi/handle/10024/32275
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):
Julkunen, Henna. “Käyttöohje voilinjan CIP-pesuprosessista
.” 2011. Thesis, Theseus. Accessed January 22, 2020.
http://www.theseus.fi/handle/10024/32275.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Julkunen, Henna. “Käyttöohje voilinjan CIP-pesuprosessista
.” 2011. Web. 22 Jan 2020.
Vancouver:
Julkunen H. Käyttöohje voilinjan CIP-pesuprosessista
. [Internet] [Thesis]. Theseus; 2011. [cited 2020 Jan 22].
Available from: http://www.theseus.fi/handle/10024/32275.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Julkunen H. Käyttöohje voilinjan CIP-pesuprosessista
. [Thesis]. Theseus; 2011. Available from: http://www.theseus.fi/handle/10024/32275
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Queens University
18.
Usprech, Jenna.
A Therapeutic Dose of Adenosine Triphosphate (ATP) for Cartilage Tissue Engineering
.
Degree: Mechanical and Materials Engineering, 2010, Queens University
URL: http://hdl.handle.net/1974/6067
► Tissue engineering holds great promise for developing functional tissue to repair damaged articular cartilage. However, cartilaginous tissues formed in vitro typically possess poor mechanical properties…
(more)
▼ Tissue engineering holds great promise for developing functional tissue to repair damaged articular cartilage. However, cartilaginous tissues formed in vitro typically possess poor mechanical properties in comparison to native tissue due to the inability of chondrocytes to accumulate adequate amounts of extracellular matrix (ECM). While mechanical stimuli can enhance the properties of engineered cartilage, it may be more efficient to harness the underlying mechanotransduction pathways responsible. Substantial evidence suggests that the mechanotransduction signaling cascade is initiated by rapid adenosine 5’-triphosphate (ATP) release from chondrocytes (purinergic receptor pathway). Thus, the purpose of this study was to investigate the effects of exogenous ATP supplementation on the biochemical and mechanical properties of tissue engineered cartilage.
Primary bovine articular chondrocytes, seeded on MilliporeTM filters, were grown in the presence of 0, 62.5 or 250 µM ATP for a period of four weeks. Both anabolic and catabolic effects were examined and a therapeutic dose range of ATP was determined.
ATP stimulation (62.5 - 250 µM) enhanced ECM synthesis by 23 - 43% and long-term collagen accumulation by 16 - 26%, in a dose-dependent manner; however, long-term proteoglycan accumulation decreased as a result of 250 µM ATP. In addition, ATP supplementation significantly improved the mechanical properties of the developed tissues (5- to 6.5-fold increase in tissue stiffness). Interestingly, high doses of ATP (250 µM) also elicited a 2-fold increase in MMP-13 gene expression, 39% increase in MMP-13 activity, and 54% more extracellular inorganic pyrophosphate (ePPi) – an ATP degradation product. Dose-dependent increases in MMP-13 activity suggested that catabolic effects were occurring alongside anabolic effects, which initiated the investigation of a therapeutic dose of ATP. Doses of 31.25 µM and 125 µM ATP were added to cartilaginous tissues and investigated in terms of MMP-13 activity and ECM synthesis. Tissues supplemented with 62.5 – 125 µM ATP exhibited a balance between anabolic and catabolic effects.
By harnessing the purinergic receptor pathway, anabolic effects of mechanical stimuli were achieved in the absence of externally applied forces. Understanding how the catabolic effects of ATP are manifested would be valuable in order to further maximize the therapeutic potential of ATP stimulation.
Subjects/Keywords: Tissue Engineering;
ATP;
Articular Cartilage
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APA ·
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APA (6th Edition):
Usprech, J. (2010). A Therapeutic Dose of Adenosine Triphosphate (ATP) for Cartilage Tissue Engineering
. (Thesis). Queens University. Retrieved from http://hdl.handle.net/1974/6067
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):
Usprech, Jenna. “A Therapeutic Dose of Adenosine Triphosphate (ATP) for Cartilage Tissue Engineering
.” 2010. Thesis, Queens University. Accessed January 22, 2020.
http://hdl.handle.net/1974/6067.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Usprech, Jenna. “A Therapeutic Dose of Adenosine Triphosphate (ATP) for Cartilage Tissue Engineering
.” 2010. Web. 22 Jan 2020.
Vancouver:
Usprech J. A Therapeutic Dose of Adenosine Triphosphate (ATP) for Cartilage Tissue Engineering
. [Internet] [Thesis]. Queens University; 2010. [cited 2020 Jan 22].
Available from: http://hdl.handle.net/1974/6067.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Usprech J. A Therapeutic Dose of Adenosine Triphosphate (ATP) for Cartilage Tissue Engineering
. [Thesis]. Queens University; 2010. Available from: http://hdl.handle.net/1974/6067
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Texas A&M University
19.
Burkeen, Jeffrey Franklin.
Calcium Signaling Mechanisms Mediate Clock-Controlled ATP Gliotransmission among Immortalized Rat SCN2.2 Cell Cultures.
Degree: 2010, Texas A&M University
URL: http://hdl.handle.net/1969.1/ETD-TAMU-2009-08-2938
► The hypothalamus is an integral part of the brain's regulation of mammalian physiology and behavior. Among many functions, this regulatory center activates the sympathetic nervous…
(more)
▼ The hypothalamus is an integral part of the brain's regulation of mammalian physiology and behavior. Among many functions, this regulatory center activates the sympathetic nervous system, maintains appropriate body temperature, controls food intake, and controls release of hormones from the pituitary gland. Deep within the hypothalamus lie a paired cluster of cells, the suprachiasmatic nuclei (SCN), which function as the chief circadian pacemaker. The goal of the present thesis research was to study rhythmically controlled
ATP gliotransmission. I used an immortalized SCN2.2 hypothalamic cell line to determine the mechanism by which
ATP signaling is regulated in this context. Additionally, this research aimed to elucidate if clock-controlled
ATP gliotransmission is fundamentally distinct from stimulus-evoked calcium-dependent mechanisms that regulate intercellular
ATP signaling among astrocytes.
In this thesis, I show that there are multiple
ATP signaling mechanisms present among SCN2.2 cells. cAMP-dependent signaling mediates clock-controlled
ATP accumulation but not stimulus-evoked
ATP signaling. In addition, pharmacological studies suggest that disparate purinergic receptor-mediated mechanisms are involved in the regulation of clock-controlled versus stimulus-evoked
ATP signaling.
Rhythmic accumulation of
ATP in SCN2.2 cultures is modulated by calcium-dependent processes. Peaks in
ATP accumulation coincide with elevated mitochondrial calcium levels, while troughs in
ATP accumulation coincide with periods of high cytosolic calcium levels, suggesting a possible mechanistic link between circadian shifts in intracellular calcium handling and
ATP handling in SCN2.2 cells. Clock-controlled
ATP accumulation in SCN2.2 cells is not a by-product of rhythmic cell cycle or rhythmic cell death.
Overall, my research suggests that the
ATP accumulation rhythm in SCN2.2 cells is likely an output of the biological clock, mediated by astrocytic calcium signaling processes, and not an output of cell division or cell death. Estimation of
ATP accumulation in SCN2.2 cultures at peak time points suggests that clock-controlled
ATP release is critical to the function of astrocytes in the mammalian brain, perhaps in the regulation of brain metabolism, the regulation of sleep/wake physiology, or the integration of both.
Advisors/Committee Members: Zoran, Mark J. (advisor), Earnest, David J. (committee member), Burghardt, Robert C. (committee member).
Subjects/Keywords: ATP; Clock-controlled ATP gliotransmission; Calcium; Calcium signaling mechanisms
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APA ·
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APA (6th Edition):
Burkeen, J. F. (2010). Calcium Signaling Mechanisms Mediate Clock-Controlled ATP Gliotransmission among Immortalized Rat SCN2.2 Cell Cultures. (Thesis). Texas A&M University. Retrieved from http://hdl.handle.net/1969.1/ETD-TAMU-2009-08-2938
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):
Burkeen, Jeffrey Franklin. “Calcium Signaling Mechanisms Mediate Clock-Controlled ATP Gliotransmission among Immortalized Rat SCN2.2 Cell Cultures.” 2010. Thesis, Texas A&M University. Accessed January 22, 2020.
http://hdl.handle.net/1969.1/ETD-TAMU-2009-08-2938.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Burkeen, Jeffrey Franklin. “Calcium Signaling Mechanisms Mediate Clock-Controlled ATP Gliotransmission among Immortalized Rat SCN2.2 Cell Cultures.” 2010. Web. 22 Jan 2020.
Vancouver:
Burkeen JF. Calcium Signaling Mechanisms Mediate Clock-Controlled ATP Gliotransmission among Immortalized Rat SCN2.2 Cell Cultures. [Internet] [Thesis]. Texas A&M University; 2010. [cited 2020 Jan 22].
Available from: http://hdl.handle.net/1969.1/ETD-TAMU-2009-08-2938.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Burkeen JF. Calcium Signaling Mechanisms Mediate Clock-Controlled ATP Gliotransmission among Immortalized Rat SCN2.2 Cell Cultures. [Thesis]. Texas A&M University; 2010. Available from: http://hdl.handle.net/1969.1/ETD-TAMU-2009-08-2938
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
20.
Rita Gabriela Pedrosa Ribeiro Mendes.
Identificação de alvos antigênicos na ATP difosfohidrolase solúvel de Schistosoma mansoni e possível aplicação funcional de peptídeos sintéticos.
Degree: 2010, Universidade Federal de Juiz de Fora
URL: http://www.bdtd.ufjf.br/tde_busca/arquivo.php?codArquivo=921
► The antigenicity of both soluble S. mansoni ATP diphosphohydrolase and membraneassociated isoforms has been recently described. Cross-immunoreactivity between potato apyrase and IgG antibody from S.…
(more)
▼ The antigenicity of both soluble S. mansoni
ATP diphosphohydrolase and membraneassociated isoforms has been recently described. Cross-immunoreactivity between potato apyrase and IgG antibody from S. mansoni-infected mice or schistosomiasis patients strongly suggested that the epitopes shared between the vegetable and parasite proteins are antigenic. By bioinformatics analysis, a strategic comparative study of the conserved domains shared between the soluble
ATP diphosphohydrolase isoform and potato apyrase predicted a homologue antigenic-domain available for antibody and HLA-DR binding. Synthetic peptides belonging to the domain of the parasite (Sm) and vegetable (pot) proteins were then obtained. By ELISA, the serum samples from the schistosomiasis patients (n= 146) showed high seropositivity of IgG1 (58%) and IgG4 (58%) antibody sub-classes for potato apyrase. Seropositive patients (n= 35) were then selected for analyses of antibody reactivity against the synthetic peptides. Significant seropositivity of IgG (potB1LJ, 9%; potB1MP, 54%; potB2LJ, 54%; SmB1LJ, 23%; SmB1MP, 86%; SmB2LJ, 34%), and IgG1 (potB1LJ, 57%; potB1MP, 67%; potB2LJ, 46%; SmB1LJ, 57%; SmB1MP, 10%; SmB2LJ, 31%) or IgG4 (potB1LJ, 71%; potB1MP, 40%; potB2LJ, 71%; SmB1LJ, 54%; SmB1MP, 63%; SmB2LJ, 49%) was observed for the homologue peptides in accordance with the existence of shared antigenic epitopes between the parasite and vegetable proteins, and demonstrating that in schistosomiasis the domain r155-194 from soluble
ATP diphosphohydrolase isoform is rich in B-cell epitopes. The reactivity of the IgG1 and IgG4 antibody from the serum samples of patients (n= 26) cured after chemotherapy with praziquantel was analyzed against potB1LJ e potB2LJ, before and six months after treatment. Individual analysis revealed variable IgG1 and IgG4 reactivity against each peptide, suggesting that different antigenic epitopes could be involved in susceptibility or resistance to S. mansoni reinfection. By ELISA, polyclonal antibody to potato apyrase of BALB/c or CBA/J mice reacts with potB1LJ, potB1MP and/or potB2LJ. Similar reactivity was also observed between these immune sera and SmB1LJ, SmB1MP and/or SmB2LJ, confirming again the cross-immunoreactivity between parasite and vegetable proteins. In addition, inoculation of each peptide in healthy Swiss mice had significant immunostimulatory activity, increasing the amount of total IgG and/or IgG1 and IgG2a antibody, as observed by ELISA and/or Dot blots, suggesting that each peptide has two distinct epitopes capable of inducing a Th1 or Th2-type immune response. By Western blots, serum immune anti-SmB2LJ recognized two polypeptides of approximately 63 and 55 kDa in SEA or SWAP, indicating the presence of the soluble isoform of
ATP diphosphohydrolase in these antigenic preparations, as well as the sensitivity and specificity of this immune serum. The potato apyrase and potB1LJ were evaluated in delayed-type hypersensitivity reaction (DTH), using ovalbumin (OVA) as standard antigen, by footpad thickness measurement,…
Advisors/Committee Members: Eveline Gomes Vasconcelos, José Otávio do Amaral Correa, Florence Mara Rosa.
Subjects/Keywords: Schistosoma mansoni; Esquistossomose; Atp Difosfohidrolase; Schistosoma mansoni; schistosomiasis; ATP diphosphohydrolase; IMUNOLOGIA
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APA (6th Edition):
Mendes, R. G. P. R. (2010). Identificação de alvos antigênicos na ATP difosfohidrolase solúvel de Schistosoma mansoni e possível aplicação funcional de peptídeos sintéticos. (Thesis). Universidade Federal de Juiz de Fora. Retrieved from http://www.bdtd.ufjf.br/tde_busca/arquivo.php?codArquivo=921
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):
Mendes, Rita Gabriela Pedrosa Ribeiro. “Identificação de alvos antigênicos na ATP difosfohidrolase solúvel de Schistosoma mansoni e possível aplicação funcional de peptídeos sintéticos.” 2010. Thesis, Universidade Federal de Juiz de Fora. Accessed January 22, 2020.
http://www.bdtd.ufjf.br/tde_busca/arquivo.php?codArquivo=921.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Mendes, Rita Gabriela Pedrosa Ribeiro. “Identificação de alvos antigênicos na ATP difosfohidrolase solúvel de Schistosoma mansoni e possível aplicação funcional de peptídeos sintéticos.” 2010. Web. 22 Jan 2020.
Vancouver:
Mendes RGPR. Identificação de alvos antigênicos na ATP difosfohidrolase solúvel de Schistosoma mansoni e possível aplicação funcional de peptídeos sintéticos. [Internet] [Thesis]. Universidade Federal de Juiz de Fora; 2010. [cited 2020 Jan 22].
Available from: http://www.bdtd.ufjf.br/tde_busca/arquivo.php?codArquivo=921.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Mendes RGPR. Identificação de alvos antigênicos na ATP difosfohidrolase solúvel de Schistosoma mansoni e possível aplicação funcional de peptídeos sintéticos. [Thesis]. Universidade Federal de Juiz de Fora; 2010. Available from: http://www.bdtd.ufjf.br/tde_busca/arquivo.php?codArquivo=921
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Vanderbilt University
21.
Scism, Robert Allen.
Directed evolution and pathway engineering for nucleotide analogue biosynthesis.
Degree: PhD, Chemistry, 2010, Vanderbilt University
URL: http://etd.library.vanderbilt.edu//available/etd-05112010-084149/
;
► Nucleotide analogues are essential pharmaceuticals with challenges in preparation due to need for control over regio and stereochemistry, and protecting group manipulations. Enzymatic biosynthesis of…
(more)
▼ Nucleotide analogues are essential pharmaceuticals with challenges in preparation due to need for control over regio and stereochemistry, and protecting group manipulations. Enzymatic biosynthesis of these analogues provides solutions to these synthetic issues, but suffers from stringent substrate requirements of the enzymes. Directed evolution presents a possible solution to amend these enzymes toward alternative substrate acceptance. We propose a superior method for nucleotide analogue production, achieved through directed evolution of nucleotide synthesizing enzymes, using an in vivo screening method. Combination of the evolved biocatalyst into an engineered biosynthetic pathway will provide an efficient system for nucleotide analogue production from ribose.
This dissertation describes using directed evolution as a means to alter whole cell turnover rates and active site specificity of hypoxanthine phosphoribosyl transferase (HPRT) for increased production of the nucleotide analogue ribavirin monophosphate. A novel in vivo screening method for negative selection of variant HPRT enzymes is introduced, and the subsequent validation of the screening method and characterization of the mutant encoded enzymes is described.
The utilization of the evolved HPRT as a general nucleotide analogue biocatalyst is detailed. A method for assaying general nucleobase turnover, and end product purification and characterization is also described. Moreover, the addition of a series of enzymes which allow a multistep synthesis of nucleotides from ribose is illustrated. Inclusion of additional enzymes for cofactor recycling adds utility and relieves byproduct inhibition. Finally, stability and reusability of the pathway is increased by self-immobilization in a cross-linked enzyme aggregate.
The sum of this work presents a significant improvement over current methods in nucleotide synthesis by alleviating current impediments due to lack of control over regio and stereochemistry, and by eliminating need for isolation of unstable intermediates, thus providing a green synthetic alternative in the synthesis of nucleotide analogues.
Advisors/Committee Members: David Wright (committee member), Richard Armstrong (committee member), Lawrence Marnett (committee member), Brian Bachmann (chair), Michael Stone (committee member).
Subjects/Keywords: atp regeneration; atp recycling; CLEA; immobilized; glutaraldehyde; biocatalysis
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APA ·
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APA (6th Edition):
Scism, R. A. (2010). Directed evolution and pathway engineering for nucleotide analogue biosynthesis. (Doctoral Dissertation). Vanderbilt University. Retrieved from http://etd.library.vanderbilt.edu//available/etd-05112010-084149/ ;
Chicago Manual of Style (16th Edition):
Scism, Robert Allen. “Directed evolution and pathway engineering for nucleotide analogue biosynthesis.” 2010. Doctoral Dissertation, Vanderbilt University. Accessed January 22, 2020.
http://etd.library.vanderbilt.edu//available/etd-05112010-084149/ ;.
MLA Handbook (7th Edition):
Scism, Robert Allen. “Directed evolution and pathway engineering for nucleotide analogue biosynthesis.” 2010. Web. 22 Jan 2020.
Vancouver:
Scism RA. Directed evolution and pathway engineering for nucleotide analogue biosynthesis. [Internet] [Doctoral dissertation]. Vanderbilt University; 2010. [cited 2020 Jan 22].
Available from: http://etd.library.vanderbilt.edu//available/etd-05112010-084149/ ;.
Council of Science Editors:
Scism RA. Directed evolution and pathway engineering for nucleotide analogue biosynthesis. [Doctoral Dissertation]. Vanderbilt University; 2010. Available from: http://etd.library.vanderbilt.edu//available/etd-05112010-084149/ ;
22.
Kabala, Anna Magdalena.
Biogenesis of mitochondrial ATP synthase and its dysfunction leading to diseases : Biogenese de l’ATP synthase mitochondriale et des dysfonctions générant des maladies.
Degree: Docteur es, Biochimie, 2014, Bordeaux; ACADEMIE POLONAISE DES SCIENCES
URL: http://www.theses.fr/2014BORD0366
► La F1FO-ATP synthase mitochondriale produit la majorité de l’énergie cellulaire chezles eucaryotes aérobes sous forme d’ATP par le processus des oxydations phosphorylantes.Chez la plupart des…
(more)
▼ La F1FO-ATP synthase mitochondriale produit la majorité de l’énergie cellulaire chezles eucaryotes aérobes sous forme d’ATP par le processus des oxydations phosphorylantes.Chez la plupart des espèces, cette enzyme possède une origine génétique double, nucléaire etmitochondriale. Dans la première partie de ce travail, je décris la construction de modèles delevure de mutations du gène mitochondrial ATP6 de l’ATP synthase découvertes chez despatients atteints de maladies neurologiques (9185T>C and 9191T>C) ou dans des tumeurs(8716A>G, 8914C>A, 8932C>T, 8953A>G and 9131T>C). Le gène ATP6 code une sousunitéessentielle (a/6) du domaine FO de l’ATP synthase. J’ai trouvé que la mutation 9185T>Cn’affecte pas l’assemblage de l’ATP synthase, mais conduit à une diminution de la vitesse desynthèse d’ATP d’environ 30%. La mutation 9191T>C empêche presque entièrementl’incorporation de la sous-unité a/6 dans l’ATP synthase. Les cinq mutations identifiées dansles tumeurs ont un effet modeste à nul, indiquant que ces mutations ne favorisent pas latumorigenèse en affectant le processus énergétique mitochondrial, comme évoquéprécédemment. J’ai ensuite étudié la régulation de la synthèse des sous-unités a/6 et 9 dans lesmitochondries de levures. La sous-unité 9 est présente sous la forme d’un anneau de 10 copiesqui interagit avec la sous-unité 6. Durant la catalyse, la rotation de cet anneau provoque deschangements conformationnels favorisant la synthèse d’ATP dans le secteur F1 de l’ATPsynthase. Je montre que la synthèse de ces protéines est couplée à leur assemblage, demanière à ce qu’elles soient produites dans une stoechiométrie adéquate et pour éviterl’accumulation d’intermédiaires d’ATP synthase potentiellement délétères
Mitochondrial F1FO-ATP synthase produces most of the cellular energy in aerobiceukaryotes under the form of ATP in the process of oxidative phosphorylation. This enzymehas in most species a double genetic origin, nuclear and mitochondrial. In the first part of thiswork, I describe the construction of yeast models of ATP synthase mutations in themitochondrial ATP6 gene, that have been found in patients presenting with neurologicaldisorders (9185T>C and 9191T>C) and in tumors (8716A>G, 8914C>A, 8932C>T,8953A>G and 9131T>C). The ATP6 gene encodes an essential subunit (called a/6) of theATP synthase proton-translocating domain (FO). The 9185T>C mutation had no effect on theassembly of ATP synthase, but reduces the rate of ATP synthesis by 30%. The 9191T>Cmutation almost completely prevented incorporation of the subunit a/6 into the ATP synthase.The five mutations found in tumors had modest, if at all, effect, indicating that thesemutations probably do not favor tumorigenesis, as was hypothesized. In the second part of mythesis, I studied the regulation of synthesis of subunits a/6 and 9 in yeast mitochondria. Thesubunit 9 is present in 10 copies forming a ring that interacts with subunit 6. Protonmovements through the FO induce the rotation of the subunit 9-ring, which results inconformational changes that promote…
Advisors/Committee Members: Di Rago, Jean-Paul (thesis director), Kucharczyk, Roza (thesis director).
Subjects/Keywords: ATP synthase; Syndrome NARP; Cancer; Biogenese de l’ATP synthase; ATP synthase; NARP syndrome; Cancer; Biogenesis of ATP synthase
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APA (6th Edition):
Kabala, A. M. (2014). Biogenesis of mitochondrial ATP synthase and its dysfunction leading to diseases : Biogenese de l’ATP synthase mitochondriale et des dysfonctions générant des maladies. (Doctoral Dissertation). Bordeaux; ACADEMIE POLONAISE DES SCIENCES. Retrieved from http://www.theses.fr/2014BORD0366
Chicago Manual of Style (16th Edition):
Kabala, Anna Magdalena. “Biogenesis of mitochondrial ATP synthase and its dysfunction leading to diseases : Biogenese de l’ATP synthase mitochondriale et des dysfonctions générant des maladies.” 2014. Doctoral Dissertation, Bordeaux; ACADEMIE POLONAISE DES SCIENCES. Accessed January 22, 2020.
http://www.theses.fr/2014BORD0366.
MLA Handbook (7th Edition):
Kabala, Anna Magdalena. “Biogenesis of mitochondrial ATP synthase and its dysfunction leading to diseases : Biogenese de l’ATP synthase mitochondriale et des dysfonctions générant des maladies.” 2014. Web. 22 Jan 2020.
Vancouver:
Kabala AM. Biogenesis of mitochondrial ATP synthase and its dysfunction leading to diseases : Biogenese de l’ATP synthase mitochondriale et des dysfonctions générant des maladies. [Internet] [Doctoral dissertation]. Bordeaux; ACADEMIE POLONAISE DES SCIENCES; 2014. [cited 2020 Jan 22].
Available from: http://www.theses.fr/2014BORD0366.
Council of Science Editors:
Kabala AM. Biogenesis of mitochondrial ATP synthase and its dysfunction leading to diseases : Biogenese de l’ATP synthase mitochondriale et des dysfonctions générant des maladies. [Doctoral Dissertation]. Bordeaux; ACADEMIE POLONAISE DES SCIENCES; 2014. Available from: http://www.theses.fr/2014BORD0366
23.
Martins, Rodrigo Machado.
A transposição didática do papel termodinâmico do ATP gera conceitos alternativos?.
Degree: Mestrado, Bioquímica, 2012, University of São Paulo
URL: http://www.teses.usp.br/teses/disponiveis/46/46131/tde-26042013-092432/
;
► Conceitos prévios, ou alternativos, são ideias introjetadas no universo cognitivo dos estudantes que diferem daqueles credenciados pela ciência estabelecida. São bastante estudadas as consequências da…
(more)
▼ Conceitos prévios, ou alternativos, são ideias introjetadas no universo cognitivo dos estudantes que diferem daqueles credenciados pela ciência estabelecida. São bastante estudadas as consequências da presença deste tipo de conceitos para o aprendizado. Entre outras dificuldades provocadas pelos conceitos alternativos está a impossibilidade de utilizá-los para embasar novos conhecimentos. Nestas condições, incapazes de decodificar apropriadamente as novas informações apresentadas pelas disciplinas, os alunos são inconscientemente encaminhados para a memorização. A origem dos conceitos prévios é variada. Uma das causas já detectadas é a precariedade da transposição didática, feita pelos autores dos livros textos ou por docentes. Uma das moléculas fundamentais para os estudos bioquímicos é a adenosina trifosfato (ATP), que desempenha múltiplas funções. Uma das principais é participar de processos que requerem energia. A compreensão do papel desta molécula é fundamental para o entendimento dos processos dos quais ela participa. Um dos objetivos do presente trabalho foi investigar equívocos sobre o papel termodinâmico do ATP nos processos celulares. Os testes foram realizados com alunos do ensino médio (EM), graduação (G) e pós-graduação (PG). A existência de concepções alternativas foi verificada, assim como sua estabilidade nos diferentes níveis de escolaridade: um resultado mostra que 68% EM, 92% G e 91% PG afirmaram que a energia da hidrólise de ATP é responsável por conduzir os processos celulares. Os resultados gerais mostram que os estudantes carregam equívocos em conceitos termodinâmicos básicos, tais como transferência de energia e espontaneidade de reações químicas. Duas possíveis fontes de conceitos alternativos da termodinâmica do ATP são o professor e o livro didático. Nesse trabalho foi verificado que os livros de ensino médio e graduação podem contribuir para a instalação de conceitos alternativos referentes ao ATP. Nos livros analisados, principalmente os de ensino médio, foram encontrados passagens, analogias e esquemas que podem contribuir para isso. E por fim, uma interferência didática foi feita com o intuito de corrigir o entendimento dos alunos no que diz respeito ao papel do ATP nas reações químicas. Alunos monitorados por meio de pré e pós-testes apresentaram resultados animadores em relação à atenuação de conceitos relacionados à termodinâmica do ATP. Dos alunos que participaram da intervenção, mais de 80% responderam e justificaram corretamente os testes feitos pós-intervenção
Misconceptions, or alternative concepts, are introjected ideas in the students` cognitive universe that differ from those established by science. The consequences of alternative concepts for the learning process are widely studied. Among other difficulties caused by alternative concepts is the impossibility to use them to support new knowledge. Under these conditions, unable to properly decode the new information submitted by the disciplines, students are unconsciously driven to memorization. The origin of the…
Advisors/Committee Members: Torres, Bayardo Baptista.
Subjects/Keywords: Alternative concepts about ATP; Biochemistry (Study and teaching); Bioquímica (Estudo e ensino); Conceitos alternativos sobre o ATP; Didactic transposition (ATP); Termodinâmica (Conceitos alternativos); Thermodynamic (Alternative concepts); Transposição (Didática ATP)
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APA (6th Edition):
Martins, R. M. (2012). A transposição didática do papel termodinâmico do ATP gera conceitos alternativos?. (Masters Thesis). University of São Paulo. Retrieved from http://www.teses.usp.br/teses/disponiveis/46/46131/tde-26042013-092432/ ;
Chicago Manual of Style (16th Edition):
Martins, Rodrigo Machado. “A transposição didática do papel termodinâmico do ATP gera conceitos alternativos?.” 2012. Masters Thesis, University of São Paulo. Accessed January 22, 2020.
http://www.teses.usp.br/teses/disponiveis/46/46131/tde-26042013-092432/ ;.
MLA Handbook (7th Edition):
Martins, Rodrigo Machado. “A transposição didática do papel termodinâmico do ATP gera conceitos alternativos?.” 2012. Web. 22 Jan 2020.
Vancouver:
Martins RM. A transposição didática do papel termodinâmico do ATP gera conceitos alternativos?. [Internet] [Masters thesis]. University of São Paulo; 2012. [cited 2020 Jan 22].
Available from: http://www.teses.usp.br/teses/disponiveis/46/46131/tde-26042013-092432/ ;.
Council of Science Editors:
Martins RM. A transposição didática do papel termodinâmico do ATP gera conceitos alternativos?. [Masters Thesis]. University of São Paulo; 2012. Available from: http://www.teses.usp.br/teses/disponiveis/46/46131/tde-26042013-092432/ ;
University of Minnesota
24.
Kurth-Nelson, Zebulun Lloyd.
Mechanisms and functional consequences of glial signaling in the retina.
Degree: PhD, Neuroscience, 2009, University of Minnesota
URL: http://purl.umn.edu/53671
► Twenty years ago, glia were viewed as passive support cells for neurons. Since then, experiments have shown that glial cells have their own form of…
(more)
▼ Twenty years ago, glia were viewed as passive support cells for neurons. Since then, experiments have shown that glial cells have their own form of excitability with precise intracellular spatiotemporal dynamics, intercellular communication among themselves, a bidirectional dialog with neurons and synapses, and a key role in mediating blood flow changes in response to neuronal activity. Most of these experiments have been conducted in brain regions such as hippocampus, cortex, hypothalamus, and cerebellum. However, as work from our laboratory has shown, the mammalian retina is also an excellent preparation to study the active functions of glial cells.
Here, we describe two forms of active glial signaling in the retina. First, we tested the hypothesis that glial cells modulate synaptic activity in the retina. We measured synaptic strength by evoking excitatory postsynaptic currents (EPSCs) in ganglion cells with either light or an electrical stimulus. We then excited glial cells through several methods, including agonist ejection, photolysis of caged Ca2+, and depolarization. The amplitude of the synaptic currents was altered by some, but not all, of these glial stimuli, leaving us unable to draw a definitive conclusion as to whether glial excitation alone is sufficient to modulate synaptic transmission in the retina.
Second, we characterized spontaneous intercellular glial Ca2+ waves in the retina. Glial cell excitability takes the form of transient intracellular Ca2+ elevations. One of the first recognized active properties of glia was their ability to propagate these Ca2+ elevations from cell to cell in a wave-like pattern. In most previous experiments, glial Ca2+ waves were initiated by an experimenter-driven stimulus, raising doubts about whether these waves occurred naturally in the organism. We demonstrate here that these waves occur spontaneously both in intact tissue and in vivo, and that the rate of spontaneous wave generation increases as animals age. These spontaneous waves propagate by glial release of ATP and activation of ATP receptors on neighboring cells. Finally, spontaneous waves cause changes in blood vessel diameter. This is the first demonstration of a functional effect of spontaneous intercellular glial signaling.
These results suggest a functional role for glial cell signaling in the retina and raise the possibility that glial signaling may actively participate in the aging of the nervous system.
Subjects/Keywords: Aging; ATP; Calcium; Glia; Muller; Retina; Neuroscience
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APA (6th Edition):
Kurth-Nelson, Z. L. (2009). Mechanisms and functional consequences of glial signaling in the retina. (Doctoral Dissertation). University of Minnesota. Retrieved from http://purl.umn.edu/53671
Chicago Manual of Style (16th Edition):
Kurth-Nelson, Zebulun Lloyd. “Mechanisms and functional consequences of glial signaling in the retina.” 2009. Doctoral Dissertation, University of Minnesota. Accessed January 22, 2020.
http://purl.umn.edu/53671.
MLA Handbook (7th Edition):
Kurth-Nelson, Zebulun Lloyd. “Mechanisms and functional consequences of glial signaling in the retina.” 2009. Web. 22 Jan 2020.
Vancouver:
Kurth-Nelson ZL. Mechanisms and functional consequences of glial signaling in the retina. [Internet] [Doctoral dissertation]. University of Minnesota; 2009. [cited 2020 Jan 22].
Available from: http://purl.umn.edu/53671.
Council of Science Editors:
Kurth-Nelson ZL. Mechanisms and functional consequences of glial signaling in the retina. [Doctoral Dissertation]. University of Minnesota; 2009. Available from: http://purl.umn.edu/53671
University of Minnesota
25.
Atchison, Nicole Ann.
Nanomaterial solutions for the protection of insulin producing beta cells.
Degree: PhD, Biomedical Engineering, 2013, University of Minnesota
URL: http://purl.umn.edu/161507
► Islet transplantation is a promising treatment for type 1 diabetes. However, even with the many successes, islet transplantation has yet to reach its full potential.…
(more)
▼ Islet transplantation is a promising treatment for type 1 diabetes. However, even with the many successes, islet transplantation has yet to reach its full potential. Limited islet sources, loss of cell viability during isolation and culture, and post-transplant graft loss are a few of the issues preventing extensive use of islet transplantation. The application of biomaterial systems to alleviate some of the stresses affecting islet viability has led to improvements in isolation and transplantation outcomes, but problems persist. In this work we approach two distinct issues affecting islet viability; ischemic conditions and immunological attack post-transplant. Ischemic conditions have been linked to a loss of islet graft function and occur during organ preservation, islet isolation and culture, and after islets are transplanted. We show that liposomal delivery of adenosine triphosphate (ATP) toβ cells can limit cell death and loss of function in ischemic conditions. We demonstrate that by functionalizing liposomes with the fibronectin-mimetic peptide PR_b, delivery of liposomes to porcine islets and rat β cells is increased compared to nontargeted controls. Additionally, liposomes are shown to protect by providing both ATP and lipids to the ischemic cells. The delivery of ATP was investigated here but application of PR_b functionalized liposomes could be extended to other interesting cargos as well. The second area of investigation involves encapsulation of islets with silica nanoparticles to create a permselective barrier. Silica nanoparticles are an interesting material for encapsulation given their ability to be fine-tuned and further functionalized. We demonstrate that size-tunable, fluorescent silica nanoparticles can be assembled layer-by-layer on the surface of cells and that silica nanoparticle encapsulated islets are able to secrete insulin in response to a glucose challenge.
Subjects/Keywords: ATP; Beta cells; Liposome; Nanomaterials; Peptides
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APA (6th Edition):
Atchison, N. A. (2013). Nanomaterial solutions for the protection of insulin producing beta cells. (Doctoral Dissertation). University of Minnesota. Retrieved from http://purl.umn.edu/161507
Chicago Manual of Style (16th Edition):
Atchison, Nicole Ann. “Nanomaterial solutions for the protection of insulin producing beta cells.” 2013. Doctoral Dissertation, University of Minnesota. Accessed January 22, 2020.
http://purl.umn.edu/161507.
MLA Handbook (7th Edition):
Atchison, Nicole Ann. “Nanomaterial solutions for the protection of insulin producing beta cells.” 2013. Web. 22 Jan 2020.
Vancouver:
Atchison NA. Nanomaterial solutions for the protection of insulin producing beta cells. [Internet] [Doctoral dissertation]. University of Minnesota; 2013. [cited 2020 Jan 22].
Available from: http://purl.umn.edu/161507.
Council of Science Editors:
Atchison NA. Nanomaterial solutions for the protection of insulin producing beta cells. [Doctoral Dissertation]. University of Minnesota; 2013. Available from: http://purl.umn.edu/161507
Case Western Reserve University
26.
Cheng, Iteen.
Investigating The Mechanism Of ATP-dependent Degradation Of
A Bacterial Protein Involved In Nucleic Acid Metabolism.
Degree: PhD, Chemistry, 2015, Case Western Reserve University
URL: http://rave.ohiolink.edu/etdc/view?acc_num=case1426628408
► Lon is an ATP-dependent serine protease that degrades damaged, misfolded, and certain regulatory proteins in the cell in order to maintain proper cellular function. Lon…
(more)
▼ Lon is an
ATP-dependent serine protease that degrades
damaged, misfolded, and certain regulatory proteins in the cell in
order to maintain proper cellular function. Lon exists as a
homo-oligomer with one ATPase domain and one protease domain in
each sub unit. Truncated ELon mutants were used to determine the
functionality of the N-terminal domain. Through steady-state and
pre-steady-state conditions it was found that despite the removal
of ~30 % of the N-terminal, substrate degradation still takes
place. A physiological substrate of Esherichia coli Lon (ELon)
called lamda N (LN) was used to demonstrate that the directionality
and rate of cleavage. Using fluorescent model substrates (dansylLN
and sFRLN), as a tool, we have employed steady-state and pre-steady
state kinetic techniques to evaluate the rate of translocation and
cleavage under
ATP conditions. The results obtained from these
experiments suggest that ELon delivers the C-terminal of LN to the
active site first followed by the N-terminal. The translocation of
LN in the presence of
ATP shows an increasing in step curve; this
could be a result of changes in a conformation change of Lon as the
fluorescent substrate moves closer to the active site. After
complete translocation, cleavage of all peptide cleavage sites
takes place within a short time frame. The initial burst of ATPase
activity suggests it is correlated to the initial stage of the
translocation step. This indicates that
ATP hydrolysis occurs
before complete substrate translocation. Collectively, these
results can be taken and fit with a global explorer program to
yield a possible mechanism for ELon. Using this information we can
determine the differences it has with hLon (human Lon protease).
Previously, ADP was found to inhibit ELon activity. However, ADP
inhibition on hLon is unknown. The similarities and differences
obtained from hLon can provide information in the development of
new therapeutic approaches to cure certain bacterial diseases in
human.
Advisors/Committee Members: Lee, Irene (Advisor), Zagorski, Michael (Committee Chair).
Subjects/Keywords: Biochemistry; Chemistry; Microbiology; Lon; protease; ATP-dependent
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APA (6th Edition):
Cheng, I. (2015). Investigating The Mechanism Of ATP-dependent Degradation Of
A Bacterial Protein Involved In Nucleic Acid Metabolism. (Doctoral Dissertation). Case Western Reserve University. Retrieved from http://rave.ohiolink.edu/etdc/view?acc_num=case1426628408
Chicago Manual of Style (16th Edition):
Cheng, Iteen. “Investigating The Mechanism Of ATP-dependent Degradation Of
A Bacterial Protein Involved In Nucleic Acid Metabolism.” 2015. Doctoral Dissertation, Case Western Reserve University. Accessed January 22, 2020.
http://rave.ohiolink.edu/etdc/view?acc_num=case1426628408.
MLA Handbook (7th Edition):
Cheng, Iteen. “Investigating The Mechanism Of ATP-dependent Degradation Of
A Bacterial Protein Involved In Nucleic Acid Metabolism.” 2015. Web. 22 Jan 2020.
Vancouver:
Cheng I. Investigating The Mechanism Of ATP-dependent Degradation Of
A Bacterial Protein Involved In Nucleic Acid Metabolism. [Internet] [Doctoral dissertation]. Case Western Reserve University; 2015. [cited 2020 Jan 22].
Available from: http://rave.ohiolink.edu/etdc/view?acc_num=case1426628408.
Council of Science Editors:
Cheng I. Investigating The Mechanism Of ATP-dependent Degradation Of
A Bacterial Protein Involved In Nucleic Acid Metabolism. [Doctoral Dissertation]. Case Western Reserve University; 2015. Available from: http://rave.ohiolink.edu/etdc/view?acc_num=case1426628408
Ruhr Universität Bochum
27.
Sojka, Johann Sebastian.
Interaktion vasokonstriktiver P2X- und P2Y-Rezeptoren in
der Mäuseniere.
Degree: 2010, Ruhr Universität Bochum
URL: http://nbn-resolving.de/urn/resolver.pl?urn=urn:nbn:de:hbz:294-27646
► Das sympathische Nervensystem ist an der Regulation des renovaskulären Widerstands beteiligt. Noradrenalin und ATP vermitteln dabei als Neurotransmitter vasokonstriktive Effekte. Postsynaptische Wirkungen von Noradrenalin und…
(more)
▼ Das sympathische Nervensystem ist an der Regulation
des renovaskulären Widerstands beteiligt. Noradrenalin und
ATP
vermitteln dabei als Neurotransmitter vasokonstriktive Effekte.
Postsynaptische Wirkungen von Noradrenalin und extrazellulären
Nucleotiden, die involvierten purinergen Rezeptoren sowie ihre
Interaktionen wurden am Modell isoliert perfundierter Niere von
Wildtyp- und P2Y4-Rezeptor-Knockout-Mäusen analysiert. Renale
Nervenstimulationen lösen frequenzabhängig Druckerhöhungen und
neuronale Noradrenalinfreisetzung aus. Neuronal und nicht neuronal
freigesetzte Nucleotide rufen durch Aktivierung von P2X1,3- und
P2Y6-Rezeptoren renale Vasokonstriktionen hervor. Beide
Rezeptorsubtypen interagieren miteinander, indem die
Desensitisierung von P2X1,3-Rezeptoren, die durch P2Y6-Rezeptoren
vermittelte Vasokonstriktionen potenziert. Bei sympathischer
Überaktivität werden extrazelluläre Nucleotide freigesetzt und
können zur Entwicklung einer essentiellen Hypertonie
beitragen.
Advisors/Committee Members: Medizin.
Subjects/Keywords: ATP; Essentielle Hypertonie; Niere; Noradrenalin;
Nucleotide
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Export
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APA (6th Edition):
Sojka, J. S. (2010). Interaktion vasokonstriktiver P2X- und P2Y-Rezeptoren in
der Mäuseniere. (Thesis). Ruhr Universität Bochum. Retrieved from http://nbn-resolving.de/urn/resolver.pl?urn=urn:nbn:de:hbz:294-27646
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):
Sojka, Johann Sebastian. “Interaktion vasokonstriktiver P2X- und P2Y-Rezeptoren in
der Mäuseniere.” 2010. Thesis, Ruhr Universität Bochum. Accessed January 22, 2020.
http://nbn-resolving.de/urn/resolver.pl?urn=urn:nbn:de:hbz:294-27646.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Sojka, Johann Sebastian. “Interaktion vasokonstriktiver P2X- und P2Y-Rezeptoren in
der Mäuseniere.” 2010. Web. 22 Jan 2020.
Vancouver:
Sojka JS. Interaktion vasokonstriktiver P2X- und P2Y-Rezeptoren in
der Mäuseniere. [Internet] [Thesis]. Ruhr Universität Bochum; 2010. [cited 2020 Jan 22].
Available from: http://nbn-resolving.de/urn/resolver.pl?urn=urn:nbn:de:hbz:294-27646.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Sojka JS. Interaktion vasokonstriktiver P2X- und P2Y-Rezeptoren in
der Mäuseniere. [Thesis]. Ruhr Universität Bochum; 2010. Available from: http://nbn-resolving.de/urn/resolver.pl?urn=urn:nbn:de:hbz:294-27646
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Ruhr Universität Bochum
28.
Wagenhäuser, Peter Johannes.
Einfluß purinerger P2Y2-Rezeptoren auf die Progredienz
der chronischen Niereninsuffizienz am Mausmodell.
Degree: 2011, Ruhr Universität Bochum
URL: http://nbn-resolving.de/urn/resolver.pl?urn=urn:nbn:de:hbz:294-32936
► Methode: Bei Wildtyp- (WT-) und P2Y2-Knockout- (KO-) Mäusen wurde durch 5/6-Nephrektomie (SNx) eine Niereninsuffizienz induziert. Bei Kontrollgruppen wurden die Nieren in situ belassen (Sham). Postoperativ…
(more)
▼ Methode: Bei Wildtyp- (WT-) und P2Y2-Knockout-
(KO-) Mäusen wurde durch 5/6-Nephrektomie (SNx) eine
Niereninsuffizienz induziert. Bei Kontrollgruppen wurden die Nieren
in situ belassen (Sham). Postoperativ erfaßte klinische und
laborchemische Parameter dokumentierten den Verlauf der
Nierenfunktion. Ergebnis: Die Sham-Gruppen zeigten keine
signifikant verschiedenen Tendenzen. Die SNx-Tiere wiesen hingegen
neben einer gegenüber den Sham-Gruppen signifikant schlechteren
Nierenfunktion auch untereinander deutliche Differenzen auf: Für
die SNx-KO-Mäuse wurde anhand von zum Beispiel Serumharnstoff,
Kreatinin-Clearance und systolischem Blutdruck eine signifikant
ausgeprägtere Niereninsuffizienz als bei den SNx-WT-Mäusen
ermittelt. Diskussion: Die vorliegende Arbeit zeigt, daß dem
P2Y2-Rezeptor im Rahmen einer renalen Schädigung möglicherweise
eine protektive Rolle zukommt.
Advisors/Committee Members: Medizin.
Subjects/Keywords: Nephrektomie; Knockout (Molekulargenetik);
Nierenkrankheit; Purinozeptor; ATP
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APA ·
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MLA ·
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Manager
APA (6th Edition):
Wagenhäuser, P. J. (2011). Einfluß purinerger P2Y2-Rezeptoren auf die Progredienz
der chronischen Niereninsuffizienz am Mausmodell. (Thesis). Ruhr Universität Bochum. Retrieved from http://nbn-resolving.de/urn/resolver.pl?urn=urn:nbn:de:hbz:294-32936
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):
Wagenhäuser, Peter Johannes. “Einfluß purinerger P2Y2-Rezeptoren auf die Progredienz
der chronischen Niereninsuffizienz am Mausmodell.” 2011. Thesis, Ruhr Universität Bochum. Accessed January 22, 2020.
http://nbn-resolving.de/urn/resolver.pl?urn=urn:nbn:de:hbz:294-32936.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Wagenhäuser, Peter Johannes. “Einfluß purinerger P2Y2-Rezeptoren auf die Progredienz
der chronischen Niereninsuffizienz am Mausmodell.” 2011. Web. 22 Jan 2020.
Vancouver:
Wagenhäuser PJ. Einfluß purinerger P2Y2-Rezeptoren auf die Progredienz
der chronischen Niereninsuffizienz am Mausmodell. [Internet] [Thesis]. Ruhr Universität Bochum; 2011. [cited 2020 Jan 22].
Available from: http://nbn-resolving.de/urn/resolver.pl?urn=urn:nbn:de:hbz:294-32936.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Wagenhäuser PJ. Einfluß purinerger P2Y2-Rezeptoren auf die Progredienz
der chronischen Niereninsuffizienz am Mausmodell. [Thesis]. Ruhr Universität Bochum; 2011. Available from: http://nbn-resolving.de/urn/resolver.pl?urn=urn:nbn:de:hbz:294-32936
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
29.
이, 진삼.
ATP depletion at G2 phase blocks mitotic entry through maintaining inhibitory phosphorylation of CDK1.
Degree: 2018, Ajou University
URL: http://repository.ajou.ac.kr/handle/201003/16540
;
http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000026806
► Tumor cells have been reported to have two main energy checkpoints depending on the amount of available ATP. These checkpoints regulate the progression of tumor…
(more)
▼ Tumor cells have been reported to have two main energy checkpoints depending on the amount of available ATP. These checkpoints regulate the progression of tumor cells during the G1 phase and their progression from the G2/M phase. Thus, the cell cycle depends on the time needed to produce enough ATP to overcome the energy checkpoints. Although molecular mechanism of G1 energy checkpoint has been reported, G2 energy checkpoint has not been fully explained. Here, I have shown that ATP depletion at G2 phase blocks both mitotic entry and cyclin B nucleus translocation in multiple cell lines. In addition, G2-M transition was regulated by the level of intracellular ATP. ATP depletion maintains p-Cdk1 (Y15), which is widely known to be phosphorylated by Wee1 kinase. Moreover, ATP depletion at G2 phase induced p-Chk1 (S345). Thus, I treated Wee1 inhibitor, PD166285, during ATP depletion at G2 phase. PD166285 dramatically overcome G2 arrest by ATP depletion. In addition, I also treated DNA damage response signaling inhibitor to target Chk1. However, DDR signaling inhibitor did not overcome G2 arrest by ATP depletion. Experiment using chemical inhibition and siRNA for AMPK also revealed that ATP depletion-induced G2 arrest did not result from AMPK activation. Taken together, ATP depletion at G2 phase blocks mitotic entry through maintaining inhibitory phosphorylation of Cdk1. Moreover, I observed that some the G2 arrested cells treated with PD166285 eventually died, indicating potential therapeutic implication and Wee1 inhibition upon ATPdepleted cells.
진핵세포는 다양한 스트레스에 반응하여 세포주기를 조절하는checkpoint를 가지고 있다. 진핵세포의 세포주기에서 이용 가능한 ATP의 양에 따라 2 가지 주요 energy checkpoint를 갖는 것으로 보고되었다. 이 checkpoint는 G1 기 동안 종양 세포주기의 진행과 G2 / M transition의 진행을 조절한다. G1 energy checkpoint에서는 AMPK에 의해 cyclin E가 억제되는 molecular mechanism이 보고되었지만 G2 energy checkpoint는 아직 완전히설명되지 않았다. 본 저자는 G2 기에서 ATP 결핍이 세포주기에 어떤영향을 미치는지 알아보고자 하였다. ATP를 결핍 시키는데 사용한 약물은 glycolysis를 억제하는 2-DG와 mitochondrial Cytochrome C oxidase를 억제하는 NaN3를 사용하였다. Drugs를 처리하였을 때 ATP가 asynchronous 수준으로 떨어지는 것을 확인하였다. 그 다음 mitotic entry와 cyclin B nuclear translocation을 확인한 결과 mitotic entry와 cyclin B nuclear translocation이 차단된 것을 관찰할 수 있었다. 이러한 현상이 drugs을 wash out 해 주었을 때 정상수준으로 회복되는지 알아보고자 하였다. 그 결과 drugs를 wash out 했을 때 ATP와 mitotic index가 정상 수준으로 회복하였다. 그 다음 세포주기가 멈추는 현상이 정말로 ATP결핍에 의한 현상인지 알아보고자 또 다른 energy stress를 가해주었다. 2-DG와 mitochondrial complex I 을 억제하는 phenformin을 동시에 처리한 결과 세포주기가 정지되는 것을 확인하였다. 그 다음 glucose free media로 glucose starvation 시켰을 때에도 이러한 현상이 일어나는지 확인하였다. 그 결과 drugs 처리 때보다는 약한정도 였으나 어느정도 세포분열기로의 이행이 억제되는 것을 볼 수 있었다. 그 다음 세포내 ATP를 측정한 결과 2-DG와 phenformin을 처리하였을 때는 asynchronous 수준으로 떨어지는 것을 확인하였다. 하지만 glucose starvation 조건에서는 ATP가 조금 감소하는 정도였다. 따라서 세포내 ATP의 양에 따라 세포주기가 조절된다는 것을 알 수 있었다. 이러한 현상이 일어나는 기전으로 먼저 AMPK에 의한 것인지 알아보고자 하였다. siRNA와 inhibitor를 사용하여 AMPK를 억제하였지만 세포주기 진행억제가 극복되지 않았다. 따라서 G2 energy checkpoint에서 AMPK는 큰 영향을 미치지 않는다는 것을 확인하였다. 그 다음 DNA 손상에 의한 신호전달 경로를 확인한 결과 Chk1이 인산화 되는 것을 확인하였다. 다양한 inhibitor를 통해 Chk1의 인산화를 억제하였지만 세포주기 진행억제를 극복하지 못했다. 따라서 Chk1도 별다른 영향을 미치지 않는다는…
Advisors/Committee Members: 대학원 의생명과학과, 201524774, 이, 진삼.
Subjects/Keywords: ATP depletion; Cdk1; Wee1; Energy-checkpoint
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APA (6th Edition):
이, . (2018). ATP depletion at G2 phase blocks mitotic entry through maintaining inhibitory phosphorylation of CDK1. (Thesis). Ajou University. Retrieved from http://repository.ajou.ac.kr/handle/201003/16540 ; http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000026806
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):
이, 진삼. “ATP depletion at G2 phase blocks mitotic entry through maintaining inhibitory phosphorylation of CDK1.” 2018. Thesis, Ajou University. Accessed January 22, 2020.
http://repository.ajou.ac.kr/handle/201003/16540 ; http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000026806.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
이, 진삼. “ATP depletion at G2 phase blocks mitotic entry through maintaining inhibitory phosphorylation of CDK1.” 2018. Web. 22 Jan 2020.
Vancouver:
이 . ATP depletion at G2 phase blocks mitotic entry through maintaining inhibitory phosphorylation of CDK1. [Internet] [Thesis]. Ajou University; 2018. [cited 2020 Jan 22].
Available from: http://repository.ajou.ac.kr/handle/201003/16540 ; http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000026806.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
이 . ATP depletion at G2 phase blocks mitotic entry through maintaining inhibitory phosphorylation of CDK1. [Thesis]. Ajou University; 2018. Available from: http://repository.ajou.ac.kr/handle/201003/16540 ; http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000026806
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
McMaster University
30.
Ortiz Castro, Mary.
Characterizing the interactions of ATP and DNA with the MutL Mismatch Repair protein.
Degree: MSc, 2016, McMaster University
URL: http://hdl.handle.net/11375/20279
► The fidelity of DNA replication prevents mutations that may lead to cancer predisposition or neurodegenerative diseases. One mechanism that enhances DNA replication fidelity is DNA…
(more)
▼ The fidelity of DNA replication prevents mutations that may lead to cancer predisposition or neurodegenerative diseases. One mechanism that enhances DNA replication fidelity is DNA mismatch repair, which corrects mismatches and small insertion/deletion loops that have escaped polymerase proofreading. In all eukaryotes and most prokaryotes, MutL (a key mismatch repair protein) has an intrinsic endonuclease activity that nicks the newly synthesized strand and recruits downstream factors to remove and correct errors. It has been proposed that ATP binding promotes a series of conformational changes that induce structural order within MutL and stimulates its endonuclease activity. The C-terminal domain of MutL, which harbors the endonuclease site, does not bind to DNA. This has prevented the molecular characterization of its endonuclease activity. In this thesis, we first show that MutL in B. subtilis exhibits asymmetric conformations similar to yeast and human MutL homologs. We also devise a novel approach to bypass the binding defect of the C-terminal domain by using fusion proteins. We find that these fusions bind to DNA specifically and, in the presence of the processivity clamp, can nick DNA. One of these fusion proteins in particular stimulates the nicking activity much more efficiently than the C-terminal domain alone. This work lays the foundation for the mechanistic characterization of the MutL endonuclease and provides a method to stabilize transient protein-DNA interactions.
Thesis
Master of Science (MSc)
Advisors/Committee Members: Guarné, Alba, Biochemistry and Biomedical Sciences.
Subjects/Keywords: MutL; mismatch repair; ATP; DNA; endonuclease activity
Record Details
Similar Records
Cite
Share »
Record Details
Similar Records
Cite
« Share
❌
APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
Ortiz Castro, M. (2016). Characterizing the interactions of ATP and DNA with the MutL Mismatch Repair protein. (Masters Thesis). McMaster University. Retrieved from http://hdl.handle.net/11375/20279
Chicago Manual of Style (16th Edition):
Ortiz Castro, Mary. “Characterizing the interactions of ATP and DNA with the MutL Mismatch Repair protein.” 2016. Masters Thesis, McMaster University. Accessed January 22, 2020.
http://hdl.handle.net/11375/20279.
MLA Handbook (7th Edition):
Ortiz Castro, Mary. “Characterizing the interactions of ATP and DNA with the MutL Mismatch Repair protein.” 2016. Web. 22 Jan 2020.
Vancouver:
Ortiz Castro M. Characterizing the interactions of ATP and DNA with the MutL Mismatch Repair protein. [Internet] [Masters thesis]. McMaster University; 2016. [cited 2020 Jan 22].
Available from: http://hdl.handle.net/11375/20279.
Council of Science Editors:
Ortiz Castro M. Characterizing the interactions of ATP and DNA with the MutL Mismatch Repair protein. [Masters Thesis]. McMaster University; 2016. Available from: http://hdl.handle.net/11375/20279
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