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Carnegie Mellon University
1.
Dykstra, Kaitlyn M.
Yip1A structures the mammalian endoplasmic reticulum.
Degree: 2012, Carnegie Mellon University
URL: http://repository.cmu.edu/dissertations/140
► The mammalian endoplasmic reticulum (ER) is the largest organelle in the cell, extending from the nuclear envelope throughout the cell periphery. The ER houses a…
(more)
▼ The mammalian endoplasmic reticulum (ER) is the largest organelle in the cell, extending from the nuclear envelope throughout the cell periphery. The ER houses a wide variety of vital cell processes within a single membrane bound organelle. In order to accommodate these functions and respond to the demands of the cell, the ER is partitioned into dynamically regulated subdomains, each with its own distinct structure. Despite the likely importance of ER structure for its functions, few proteins have been identified as having a direct role in maintaining the structure of the ER and the consequences of alteration of normal ER structure are not well understood.
Here we identify Yip1A, a conserved membrane protein that cycles between the ER and early Golgi, as a likely regulator of ER organization. Yip1A depletion led to restructuring of ER membranes into micrometer-sized, concentrically stacked whorls. These structures are reminiscent of the ER whorls found in certain specialized secretory cell types, where the regulation and functional consequence of ER whorl formation is not understood. We found that membrane stacking and whorl formation after Yip1A depletion coincided with a marked slowing of coat protein (COP) II-mediated protein export from the ER. Furthermore, whorl formation driven by exogenous expression of an ER protein with no role in COPII function also delayed cargo export. Thus, it appears that Yip1A is required to prevent ER whorl formation and that whorl formation can in turn delay protein export from the organelle. Whether this is the function of ER whorls in tissues remains to be seen, however these results make Yip1A a good candidate for playing a role in their regulation.
To obtain insight into how Yip1A regulates ER whorl formation and to determine whether the mechanism might be shared with the yeast homologue Yip1p, we carried out a systematic mutational analysis of all residues in the protein. Two discrete sites (E95 and K146) were crucial for the control of ER whorl formation by Yip1A. Notably, the same residues were previously shown to be important for Yip1p-mediated viability in yeast, indicating a shared mechanism. On the other hand, a third site (E89) also essential for yeast viability was dispensable for Yip1A function in regulating whorl formation. Thus Yip1p/Yip1A may possess at least two distinct essential functions only one of which is required for regulation of ER structure. Of note, the sites required for control of ER whorl formation by Yip1A were dispensable for the binding of Yip1p to its established binding partners Yif1p and Ypt1/31p, whereas the site required for Yip1p to bind the same partners was dispensable for ER structuring by Yip1A. Based on these observations, we speculate that the function of Yip1A in regulating whorl formation is mediated by one or more distinct and yet-to-be identified binding partners.
Collectively, these findings indicate that a dispersed ER network is important for proper COPII-mediated protein export and that Yip1A has a conserved function between…
Subjects/Keywords: Biochemistry; Biophysics; and Structural Biology
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APA (6th Edition):
Dykstra, K. M. (2012). Yip1A structures the mammalian endoplasmic reticulum. (Thesis). Carnegie Mellon University. Retrieved from http://repository.cmu.edu/dissertations/140
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):
Dykstra, Kaitlyn M. “Yip1A structures the mammalian endoplasmic reticulum.” 2012. Thesis, Carnegie Mellon University. Accessed March 04, 2021.
http://repository.cmu.edu/dissertations/140.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Dykstra, Kaitlyn M. “Yip1A structures the mammalian endoplasmic reticulum.” 2012. Web. 04 Mar 2021.
Vancouver:
Dykstra KM. Yip1A structures the mammalian endoplasmic reticulum. [Internet] [Thesis]. Carnegie Mellon University; 2012. [cited 2021 Mar 04].
Available from: http://repository.cmu.edu/dissertations/140.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Dykstra KM. Yip1A structures the mammalian endoplasmic reticulum. [Thesis]. Carnegie Mellon University; 2012. Available from: http://repository.cmu.edu/dissertations/140
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Virginia Commonwealth University
2.
Govinda Remesh, Soumya.
STRUCTURAL STUDIES OF INTERFERON REGULATORY FACTOR 4: A MOLECULAR PERSPECTIVE OF ITS REGULATORY MECHANISM.
Degree: PhD, Physiology and Biophysics, 2014, Virginia Commonwealth University
URL: https://doi.org/10.25772/3HWQ-5T45
;
https://scholarscompass.vcu.edu/etd/3572
► Interferon (IFN) regulatory factor family member 4 (IRF4) is a transcription factor that serves specific roles in transcriptional regulation of IFN responsive genes and…
(more)
▼ Interferon (IFN) regulatory factor family member 4 (IRF4) is a transcription factor that serves specific roles in transcriptional regulation of IFN responsive genes and is indispensable in B- & T-cell differentiation. IRF4 like the other members of the family has two major domains- the N-terminal DNA binding domain (DBD) essential for its recognition and binding to the Interferon Stimulated Response Element DNA sequence and a C-terminal Interferon activation domain (IAD) thought to maintain IRF4 in an auto-inhibited inactive state and is also critical in its activation. A putative unstructured linker connects the DBD and IAD. Activation in most members of the IRF family requires phosphorylation to induce homo and hetero-dimerization. In contrast, IRF4 functions primarily through ternary complex formation involving different proteins including PU.1 and MyD88. The IRF4IAD has a C-terminal auto-inhibitory region (AIR) that has been proposed to physically impede the DBD from interacting with DNA in the absence of its binding partner.
To understand the activation mechanism in molecular detail we determined the crystal structure of the IAD of IRF4 and also performed small-angle X-ray scattering (SAXS) studies. Our data reveals that the surface electrostatics of IAD and presence of additional loops confers exclusivity to IRF4 in the IRF family. SAXS studies suggest that the AIR is structured and makes interactions with the putative linker. We also performed analytical ultracentrifugation studies, fluorescence anisotropy binding experiments and SAXS studies on full-length IRF4 as well as on constructs where the first 20 residues, exclusive to IRF4 or the AIR were removed. We observe that the first 20 residues are critical in decreasing the binding affinity of full-length IRF4 to DNA. In addition, the putative linker of IRF4 connecting the N- and C-termini appears to be a folded domain and interacts with AIR. Also, overall full-length IRF4 appears as an elongated molecule and the N- and the C-terminal domains are arranged on either ends of full-length IRF4. Moreover, there are no signs of huge conformational changes in the protein during the activation process. Taken together, based on our data we propose that there is no auto-inhibited state for IRF4. Furthermore, it is the binding affinity of full-length IRF4 that is increased in the presence of its binding partner most likely through modest conformational changes.
Advisors/Committee Members: Dr. Carlos Escalante, Dr. Montserrat Samso, Dr. Qinglian Liu, Dr. Darrel Peterson, Dr. Xiang-Yang Wang.
Subjects/Keywords: Biochemistry; Biophysics; and Structural Biology
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❌
APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
Govinda Remesh, S. (2014). STRUCTURAL STUDIES OF INTERFERON REGULATORY FACTOR 4: A MOLECULAR PERSPECTIVE OF ITS REGULATORY MECHANISM. (Doctoral Dissertation). Virginia Commonwealth University. Retrieved from https://doi.org/10.25772/3HWQ-5T45 ; https://scholarscompass.vcu.edu/etd/3572
Chicago Manual of Style (16th Edition):
Govinda Remesh, Soumya. “STRUCTURAL STUDIES OF INTERFERON REGULATORY FACTOR 4: A MOLECULAR PERSPECTIVE OF ITS REGULATORY MECHANISM.” 2014. Doctoral Dissertation, Virginia Commonwealth University. Accessed March 04, 2021.
https://doi.org/10.25772/3HWQ-5T45 ; https://scholarscompass.vcu.edu/etd/3572.
MLA Handbook (7th Edition):
Govinda Remesh, Soumya. “STRUCTURAL STUDIES OF INTERFERON REGULATORY FACTOR 4: A MOLECULAR PERSPECTIVE OF ITS REGULATORY MECHANISM.” 2014. Web. 04 Mar 2021.
Vancouver:
Govinda Remesh S. STRUCTURAL STUDIES OF INTERFERON REGULATORY FACTOR 4: A MOLECULAR PERSPECTIVE OF ITS REGULATORY MECHANISM. [Internet] [Doctoral dissertation]. Virginia Commonwealth University; 2014. [cited 2021 Mar 04].
Available from: https://doi.org/10.25772/3HWQ-5T45 ; https://scholarscompass.vcu.edu/etd/3572.
Council of Science Editors:
Govinda Remesh S. STRUCTURAL STUDIES OF INTERFERON REGULATORY FACTOR 4: A MOLECULAR PERSPECTIVE OF ITS REGULATORY MECHANISM. [Doctoral Dissertation]. Virginia Commonwealth University; 2014. Available from: https://doi.org/10.25772/3HWQ-5T45 ; https://scholarscompass.vcu.edu/etd/3572

Iowa State University
3.
Gogerty, David.
Isobutene formation from 3-hydroxy-3-methylbutyrate (3-HMB) by the Saccharomyces cerevisiae diphosphomevalonate decarboxylase (ScMDD) and directed enzyme evolution to improve enzyme function.
Degree: 2011, Iowa State University
URL: https://lib.dr.iastate.edu/etd/10353
► Dependence on petroleum for energy and petrochemical products has led to high energy costs, a polluted environment, the depletion of a finite resource (oil), and…
(more)
▼ Dependence on petroleum for energy and petrochemical products has led to high energy costs, a polluted environment, the depletion of a finite resource (oil), and the reliance on hostile nations for our energy security. Biofuels promise to alleviate some or all of these issues but remain costly and inefficient to produce, and difficult to integrate into our existing transportation infrastructure. A renewable fuel molecule capable of replacing petroleum for our fuel and chemical industries is desired. Isobutene is an important commercial chemical used for the synthesis of butyl rubber, terephthalic acid, specialty chemicals, and a gasoline performance additive known as alkylate. Currently, isobutene is produced from petroleum and hence is nonrenewable. Here we report that the Saccharomyces cerevisiae mevalonate diphosphate decarboxylase (ScMDD) can convert 3-hydroxy-3-methylbutyrate (3-HMB) to isobutene. Whole cells of Escherichia coli producing ScMDD formed isobutene from 3-HMB at a rate of 154 pmol h-1 g cells-1. His6-ScMDD was purified by nickel affinity chromatography and shown to produce isobutene from 3-HMB at a rate of 1.33 pmol min-1 mg-1 protein. In contrast, no isobutene was detected from control cells lacking ScMDD, and controls showed that both His6-ScMDD and 3-HMB were required for detectable isobutene formation in enzyme assays. ScMDD was subjected to error-prone PCR and 2 improved variants were characterized, ScMDD1 (I145F) and ScMDD2 (R74H). Whole cells of E. coli producing ScMDD1 and ScMDD2 produced isobutene from 3-HMB at rates of 3000 and 5888 pmol h-1 g cells-1 which are 19- and 38-fold increases compared to cells producing His6-ScMDD. Although ScMDD was shown to be amenable to manipulation in order to increase its activity on 3-HMB, we estimate that a 106 fold increase in activity is needed for commercial application. Two novel methods were designed to increase enzyme activity – a mevalonate selection and an isobutene biosensor.
The mevalonate selection yielded one variant with a 3.9-fold increase in isopentenol production from mevalonate over wild-type, to 186.6 nmol min-1 g cells-1, as well as a 51.6% increase in isobutene-formation. The isobutene biosensor was built and confirmed to respond to isobutene, toluene, and isoprene with induction ratios of 1.64, 3.58, and 1.3, respectively.
Subjects/Keywords: Biochemistry; Biophysics; and Structural Biology
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❌
APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
Gogerty, D. (2011). Isobutene formation from 3-hydroxy-3-methylbutyrate (3-HMB) by the Saccharomyces cerevisiae diphosphomevalonate decarboxylase (ScMDD) and directed enzyme evolution to improve enzyme function. (Thesis). Iowa State University. Retrieved from https://lib.dr.iastate.edu/etd/10353
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):
Gogerty, David. “Isobutene formation from 3-hydroxy-3-methylbutyrate (3-HMB) by the Saccharomyces cerevisiae diphosphomevalonate decarboxylase (ScMDD) and directed enzyme evolution to improve enzyme function.” 2011. Thesis, Iowa State University. Accessed March 04, 2021.
https://lib.dr.iastate.edu/etd/10353.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Gogerty, David. “Isobutene formation from 3-hydroxy-3-methylbutyrate (3-HMB) by the Saccharomyces cerevisiae diphosphomevalonate decarboxylase (ScMDD) and directed enzyme evolution to improve enzyme function.” 2011. Web. 04 Mar 2021.
Vancouver:
Gogerty D. Isobutene formation from 3-hydroxy-3-methylbutyrate (3-HMB) by the Saccharomyces cerevisiae diphosphomevalonate decarboxylase (ScMDD) and directed enzyme evolution to improve enzyme function. [Internet] [Thesis]. Iowa State University; 2011. [cited 2021 Mar 04].
Available from: https://lib.dr.iastate.edu/etd/10353.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Gogerty D. Isobutene formation from 3-hydroxy-3-methylbutyrate (3-HMB) by the Saccharomyces cerevisiae diphosphomevalonate decarboxylase (ScMDD) and directed enzyme evolution to improve enzyme function. [Thesis]. Iowa State University; 2011. Available from: https://lib.dr.iastate.edu/etd/10353
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Iowa State University
4.
Leelananda, Sumudu Pamoda.
Protein sequence-structure relationships.
Degree: 2011, Iowa State University
URL: https://lib.dr.iastate.edu/etd/10344
► Physical characteristics of amino acids are responsible for the folding of protein sequences to their native structures. An understanding of protein sequence-structure relationships is required…
(more)
▼ Physical characteristics of amino acids are responsible for the folding of protein sequences to their native structures. An understanding of protein sequence-structure relationships is required to solve the folding problem and it is one of the most important problems in computational structural biology. Even though there are tens of thousands of protein structures in the Protein Data Bank, it is not understood why they take their particular structures or why they are limited to a few thousands of folds. It is well known that protein structures are evolutionarily more conserved than sequences and that, often, sequences with low sequence identity can share the same fold. This leads to the concept of protein designability. The designability of a particular conformation is defined as the number of different sequences that fold to it giving unique minimum energy.
Graph features of contact diagrams are employed here to describe the topology of lattice models of proteins and coarse-grained protein structures. The relationship between graphical features and designability of structures is explored here in various ways. It is found that there exists a relationship between some simple geometric graph features and designability. Highly designable structures can be distinguished from poorly designable structures based on those graphical features. This finding confirms the fact that the topology of a protein structure giving rise to its residue-residue interaction network is an important determinant of its designability.
We learn that, the higher the designability of a structure is, the more diverse is its sequence space. However, there are conserved positions, which are more frequently conserved as either polar or hydrophobic. There is a marked difference between the hydrophobic/polar profiles of highly and poorly designable sequences, and thus, they become more clearly distinguishable. These profiles can be used to train machine learning algorithms to predict the designability of sequences.
Subjects/Keywords: Biochemistry; Biophysics; and Structural Biology
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❌
APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
Leelananda, S. P. (2011). Protein sequence-structure relationships. (Thesis). Iowa State University. Retrieved from https://lib.dr.iastate.edu/etd/10344
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):
Leelananda, Sumudu Pamoda. “Protein sequence-structure relationships.” 2011. Thesis, Iowa State University. Accessed March 04, 2021.
https://lib.dr.iastate.edu/etd/10344.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Leelananda, Sumudu Pamoda. “Protein sequence-structure relationships.” 2011. Web. 04 Mar 2021.
Vancouver:
Leelananda SP. Protein sequence-structure relationships. [Internet] [Thesis]. Iowa State University; 2011. [cited 2021 Mar 04].
Available from: https://lib.dr.iastate.edu/etd/10344.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Leelananda SP. Protein sequence-structure relationships. [Thesis]. Iowa State University; 2011. Available from: https://lib.dr.iastate.edu/etd/10344
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Iowa State University
5.
Wang, Lijun.
Studies of the structure and function of Mms6, a bacterial protein that promotes the formation of magnetic nanoparticles.
Degree: 2011, Iowa State University
URL: https://lib.dr.iastate.edu/etd/12014
► Many highly ordered mineralized structures are created by living organisms that are often hierarchical in structure with fundamental structural elements at nanometer scales. The ability…
(more)
▼ Many highly ordered mineralized structures are created by living organisms that are often hierarchical in structure with fundamental structural elements at nanometer scales. The ability to fabricate such fundamental structures independently of these organisms could open many new and exciting opportunities in nanotechnology. In those cases for which there is some understanding of the biological mechanisms involved in biomineralization, proteins have been found responsible for forming the mineral structures. But, the mechanisms by which mineralization proteins function are poorly understood. Here we discuss our studies of the magnetotactic bacterial protein, Mms6, which promotes the formation in vitro of highly paramagnetic nanocrystals. Mms6 has two phases of iron binding, the first is very high affinity and the second is low affinity and cooperative with respect to iron. Our results provide evidence for a model for the mechanism of action of Mms6 in which a conformational change driven by a high affinity iron binding site in the C-terminal domain initiates a coordinated structural change involving multiple proteins to form a surface that can initiate the packing of iron into a crystal lattice. The small starting crystals might then fuse to produce larger nanoparticles as the protein islands move in a fluid hydrophobic environment of a membrane or micelles.
Subjects/Keywords: Biochemistry; Biophysics; and Structural Biology
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❌
APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
Wang, L. (2011). Studies of the structure and function of Mms6, a bacterial protein that promotes the formation of magnetic nanoparticles. (Thesis). Iowa State University. Retrieved from https://lib.dr.iastate.edu/etd/12014
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):
Wang, Lijun. “Studies of the structure and function of Mms6, a bacterial protein that promotes the formation of magnetic nanoparticles.” 2011. Thesis, Iowa State University. Accessed March 04, 2021.
https://lib.dr.iastate.edu/etd/12014.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Wang, Lijun. “Studies of the structure and function of Mms6, a bacterial protein that promotes the formation of magnetic nanoparticles.” 2011. Web. 04 Mar 2021.
Vancouver:
Wang L. Studies of the structure and function of Mms6, a bacterial protein that promotes the formation of magnetic nanoparticles. [Internet] [Thesis]. Iowa State University; 2011. [cited 2021 Mar 04].
Available from: https://lib.dr.iastate.edu/etd/12014.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Wang L. Studies of the structure and function of Mms6, a bacterial protein that promotes the formation of magnetic nanoparticles. [Thesis]. Iowa State University; 2011. Available from: https://lib.dr.iastate.edu/etd/12014
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

East Tennessee State University
6.
Yan, Hui.
Regulation of Acute and Chronic Immune Responses by β-Arrestin2.
Degree: PhD, Biomedical Sciences, 2016, East Tennessee State University
URL: https://dc.etsu.edu/etd/3049
► β-arrestin2, previously recognized as a facilitator for G-protein associated 7 TMR desensitization/ internalization, has now been appreciated as an independent signal transducer that regulates…
(more)
▼ β-arrestin2, previously recognized as a facilitator for G-protein associated 7 TMR desensitization/ internalization, has now been appreciated as an independent signal transducer that regulates multiple cellular responses including inflammation. Cecal ligation and puncture procedure (CLP) induced septic shock is an acute inflammatory response characterized by uncontrolled systemic inflammation. Myocardial ischemia/reperfusion is a chronic sterilize inflammation that requires the reaction of macrophages, fibroblasts and cardiac stem cells for regeneration and remodeling of the infarcted myocardium. Restrained chronic stress is an immune suppression model in which the inactivation of macrophages may be involved. Here we showed β-arrestin2 overexpression inhibited CLP-induced heart dysfunction in septic shock, stabilized the cardiovascular system, and eventually promoted survival. Inhibition of the activation of p38 that downstream of the IL-6 pathway may be a key regulatory target for β-arrestin2. To rescue cardiomyocytes from ischemia and reperfusion injury, Sca-1+ CSC from Wide-type or β-arrestin2 Knockout mice were delivered to the risked area before reperfusion; β-arrestin2 was shown to be a required factor and a promoter for the differentiation of the cardiac stem cells. A β-arrestin2/miR-155/GSK3β pathway was identified in this study. TLR-9 is an important part of the innate immune system which has been shown to be regulated by β-arrestin2 in various inflammatory models. Here we found, the immune suppression induced by restrained stress is mediated by Toll-like receptor 9 (TLR-9). TLR-9 facilitated the elevation of IL-1β, IL-10 and IL-17 levels in serum and decrease of the levels of plasma IFN-γ. Furthermore, macrophage apoptosis was alleviated in TLR-9 deficiency mice. In summary, β-arrestin2 and associated proteins like TLR-9 are important regulators of the immune response in a variety of disease conditions. Therapeutic strategies should be generated to balance the inflammation and anti-inflammation response by modulating β-arrestin2 expression and functions.
Subjects/Keywords: Biochemistry; Biophysics; and Structural Biology
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❌
APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
Yan, H. (2016). Regulation of Acute and Chronic Immune Responses by β-Arrestin2. (Doctoral Dissertation). East Tennessee State University. Retrieved from https://dc.etsu.edu/etd/3049
Chicago Manual of Style (16th Edition):
Yan, Hui. “Regulation of Acute and Chronic Immune Responses by β-Arrestin2.” 2016. Doctoral Dissertation, East Tennessee State University. Accessed March 04, 2021.
https://dc.etsu.edu/etd/3049.
MLA Handbook (7th Edition):
Yan, Hui. “Regulation of Acute and Chronic Immune Responses by β-Arrestin2.” 2016. Web. 04 Mar 2021.
Vancouver:
Yan H. Regulation of Acute and Chronic Immune Responses by β-Arrestin2. [Internet] [Doctoral dissertation]. East Tennessee State University; 2016. [cited 2021 Mar 04].
Available from: https://dc.etsu.edu/etd/3049.
Council of Science Editors:
Yan H. Regulation of Acute and Chronic Immune Responses by β-Arrestin2. [Doctoral Dissertation]. East Tennessee State University; 2016. Available from: https://dc.etsu.edu/etd/3049

Eastern Illinois University
7.
Gamage, Hashni Epa Vidana.
Functional Characterization of Glutamate Carboxypeptidase II in Caenorhabditis elegans.
Degree: MS, Biological Sciences, 2019, Eastern Illinois University
URL: https://thekeep.eiu.edu/theses/4571
► Glutamate carboxypeptidase II (GCPII) is a transmembrane zinc metalloprotease expressed in a number of organisms: from yeast to worm to humans. In humans, GCPII…
(more)
▼ Glutamate carboxypeptidase II (GCPII) is a transmembrane zinc metalloprotease expressed in a number of organisms: from yeast to worm to humans. In humans, GCPII has been observed as a multifunctional protein and expressed in prostate, intestine, kidney, brain, tumor-associated neovasculature and other tissues as five paralogs. In the human small intestine, hGCPII is proposed to facilitate the folate absorption by cleaving terminal glutamate residues in dietary folates. Folates act as a cofactor in one-carbon metabolic pathways such as nucleotide synthesis, amino acid synthesis, DNA repair, and consequently involved in cell division and growth. The hGCPII homolog is found in the nematode
Caenorhabditis elegans (cGCPII) as three paralogs and shares a high
structural similarity with hGCPII. In this study, the
C. elegans strains: wild-type (N2), and
gcp-2 deletion mutant strains: RB1055
(gcp-2.1), TM6632
(gcp-2.2) and TM5414
(gcp-2.3) were used to investigate the role of
gcp-2 in folate metabolism. This study shows that the
gcp-2.1 and
gcp-2.2 paralogs play a significant role in folate metabolism, reproduction, and embryonic and post-embryonic development in
C. elegans. When the
gcp-2 mutant worms were fed with a folate-deficient diet, it showed folate deficient phenotypes, infertility and growth retardation, as observed in mice and humans. This work establishes, for the first time, the relationship between GCPII and folate metabolism in
C. elegans as proposed for human folate metabolism. This study demonstrates that
C. elegans can be used as a genetically tractable model organism to invetigate the tissue-specific multifunctional roles of GCPII in development and reproduction of a multicellular organsim.
Advisors/Committee Members: Gopal R. Periyannan, Britto P. Nathan, Gary A. Bulla.
Subjects/Keywords: Biochemistry; Biophysics; and Structural Biology
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❌
APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
Gamage, H. E. V. (2019). Functional Characterization of Glutamate Carboxypeptidase II in Caenorhabditis elegans. (Masters Thesis). Eastern Illinois University. Retrieved from https://thekeep.eiu.edu/theses/4571
Chicago Manual of Style (16th Edition):
Gamage, Hashni Epa Vidana. “Functional Characterization of Glutamate Carboxypeptidase II in Caenorhabditis elegans.” 2019. Masters Thesis, Eastern Illinois University. Accessed March 04, 2021.
https://thekeep.eiu.edu/theses/4571.
MLA Handbook (7th Edition):
Gamage, Hashni Epa Vidana. “Functional Characterization of Glutamate Carboxypeptidase II in Caenorhabditis elegans.” 2019. Web. 04 Mar 2021.
Vancouver:
Gamage HEV. Functional Characterization of Glutamate Carboxypeptidase II in Caenorhabditis elegans. [Internet] [Masters thesis]. Eastern Illinois University; 2019. [cited 2021 Mar 04].
Available from: https://thekeep.eiu.edu/theses/4571.
Council of Science Editors:
Gamage HEV. Functional Characterization of Glutamate Carboxypeptidase II in Caenorhabditis elegans. [Masters Thesis]. Eastern Illinois University; 2019. Available from: https://thekeep.eiu.edu/theses/4571

Duquesne University
8.
Ferraro, Nicholas.
State-Dependent Mapping of GlyR-Cholesterol Interactions by Coupling Crosslinking with Mass Spectrometry.
Degree: PhD, Chemistry and Biochemistry, 2019, Duquesne University
URL: https://dsc.duq.edu/etd/1837
► The glycine receptor (GlyR) belongs to a superfamily of pentameric ligand-gated ion channels (pLGICs) that mediate fast neurotransmission. GlyR typically modulates inhibitory transmission by…
(more)
▼ The glycine receptor (GlyR) belongs to a superfamily of pentameric ligand-gated ion channels (pLGICs) that mediate fast neurotransmission. GlyR typically modulates inhibitory transmission by antagonizing membrane depolarization through anion influx. Allosteric interactions between the receptor and its lipid surroundings affect receptor function, and cholesterol is essential for pLGIC activity. Human α1 GlyR was purified from baculovirus infected insect cells and reconstituted in unilamellar vesicles at cholesterol: lipid ratios below and above the cholesterol activity threshold with aliquots of azi-cholesterol. State-dependent crosslinking studies of receptors primarily in its resting (no glycine), desensitized (10mM glycine) and open (F207A/A288G, 30nM ivermectin) states were then performed at elevated cholesterol levels necessary for activity. After photoactivation, covalently crosslinked cholesterol-GlyR were trypsinized, mass fingerprinted by tandem mass spectrometry (MS-MS), and sites of cholesterol crosslinks in peptides were refined by targeted MS-MS. Within the GlyR apo state, cholesterol interactions differed as a function of membrane cholesterol concentration correlating to the chemical activity of cholesterol, suggesting two distinct conformations. Differential cholesterol crosslinking patterns between resting, desensitized, and open states were observed, highlighting state-dependent differences in GlyR lipid accessibility. Distinct state-dependent crosslinking patterns indicative of alterations in either the lipid environment and/or channel structure were observed throughout GlyR, most prominently observed in the M4 transmembrane helix, extracellular domain loops and regions nearing the bilayer interface, and the large intracellular M3-M4 loop. The changes in M4 accessibility (transition from surface-mapped crosslinking to regions of the helix less exposed when mapped) suggest an outward twisting motion and translocation towards the bilayer/lipids as GlyR allosterically transitions. Strikingly, crosslinking patterns within the M3-M4 loop offer insight into the generalized structure of this unresolved region in all current pLGIC
structural models, by suggesting the crosslinked regions of this intracellular loop are intimately associated or buried within the lipid bilayer. Taken together, crosslinking coupled with MS-MS has the capability to accurately probe and define physiological protein frameworks which can aid in the refinement of allosteric modulation and current
structural models.
Advisors/Committee Members: Michael Cascio, Stephanie Wetzel, Skip Kingston, Jana Patton-Vogt.
Subjects/Keywords: Biochemistry; Biophysics; and Structural Biology
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APA (6th Edition):
Ferraro, N. (2019). State-Dependent Mapping of GlyR-Cholesterol Interactions by Coupling Crosslinking with Mass Spectrometry. (Doctoral Dissertation). Duquesne University. Retrieved from https://dsc.duq.edu/etd/1837
Chicago Manual of Style (16th Edition):
Ferraro, Nicholas. “State-Dependent Mapping of GlyR-Cholesterol Interactions by Coupling Crosslinking with Mass Spectrometry.” 2019. Doctoral Dissertation, Duquesne University. Accessed March 04, 2021.
https://dsc.duq.edu/etd/1837.
MLA Handbook (7th Edition):
Ferraro, Nicholas. “State-Dependent Mapping of GlyR-Cholesterol Interactions by Coupling Crosslinking with Mass Spectrometry.” 2019. Web. 04 Mar 2021.
Vancouver:
Ferraro N. State-Dependent Mapping of GlyR-Cholesterol Interactions by Coupling Crosslinking with Mass Spectrometry. [Internet] [Doctoral dissertation]. Duquesne University; 2019. [cited 2021 Mar 04].
Available from: https://dsc.duq.edu/etd/1837.
Council of Science Editors:
Ferraro N. State-Dependent Mapping of GlyR-Cholesterol Interactions by Coupling Crosslinking with Mass Spectrometry. [Doctoral Dissertation]. Duquesne University; 2019. Available from: https://dsc.duq.edu/etd/1837

University of Tennessee – Knoxville
9.
Bucci, Joel Cullen.
Pinpointing the Molecular Basis for Metal Ion Effects on Plasminogen Activator Inhibitor-1 (PAI-1).
Degree: 2016, University of Tennessee – Knoxville
URL: https://trace.tennessee.edu/utk_graddiss/3896
► Plasminogen activator inhibitor type-1 (PAI-1) specifically inhibits the proteases tissue type plasminogen activator and urokinase plasminogen activator to control the activation of fibrinolysis. Vitronectin interacts…
(more)
▼ Plasminogen activator inhibitor type-1 (PAI-1) specifically inhibits the proteases tissue type plasminogen activator and urokinase plasminogen activator to control the activation of fibrinolysis. Vitronectin interacts with PAI-1 primarily through the somatomedin B (SMB) domain to stabilize and localize PAI-1 to sites of injury. Our laboratory observed that transition metals such as copper2+ have VN dependent, reciprocal effects on how long PAI-1 remains active. We aim to determine the molecular basis for effects of copper2+ on PAI-1 activity. We employed a computational algorithm (MUG) to predict metal binding clusters, and introduced mutations hypothesized to create metal binding deficiency. We compared mutants to wild-type by: measurement of stability kinetics, thermodynamic parameters using isothermal titration calorimetry, and protein dynamics using hydrogen deuterium exchange. Active PAI-1 binds copper2+ in the low nanomolar range, while latent binds an order of magnitude weaker. In a mutant lacking the N-terminal histidines of PAI-1, we observed reduced copper2+ binding, but this does not abolish accelerated transition to the latent form. PAI-1 mutants lacking the carboxylate containing resides in the gate region, as well as a histidine of s4B proximal to the flexible joint region require more copper2+ than wild-type to promote accelerated latency formation, making these residues candidates for further metal binding characterization. SMB-PAI-1 complex binds copper2+ with comparable affinity and stoichiometry as PAI-1 alone. Finally, the SMB domain stabilizes PAI-1 by localized effects on dynamics in the same regions that are affected by copper2+. Thus, binding of SMB does not sterically interfere with copper binding to PAI-1, but rather negates copper2+ effects directly through changes in dynamics.
Subjects/Keywords: Biochemistry; Biophysics; Structural Biology
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APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
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APA (6th Edition):
Bucci, J. C. (2016). Pinpointing the Molecular Basis for Metal Ion Effects on Plasminogen Activator Inhibitor-1 (PAI-1). (Doctoral Dissertation). University of Tennessee – Knoxville. Retrieved from https://trace.tennessee.edu/utk_graddiss/3896
Chicago Manual of Style (16th Edition):
Bucci, Joel Cullen. “Pinpointing the Molecular Basis for Metal Ion Effects on Plasminogen Activator Inhibitor-1 (PAI-1).” 2016. Doctoral Dissertation, University of Tennessee – Knoxville. Accessed March 04, 2021.
https://trace.tennessee.edu/utk_graddiss/3896.
MLA Handbook (7th Edition):
Bucci, Joel Cullen. “Pinpointing the Molecular Basis for Metal Ion Effects on Plasminogen Activator Inhibitor-1 (PAI-1).” 2016. Web. 04 Mar 2021.
Vancouver:
Bucci JC. Pinpointing the Molecular Basis for Metal Ion Effects on Plasminogen Activator Inhibitor-1 (PAI-1). [Internet] [Doctoral dissertation]. University of Tennessee – Knoxville; 2016. [cited 2021 Mar 04].
Available from: https://trace.tennessee.edu/utk_graddiss/3896.
Council of Science Editors:
Bucci JC. Pinpointing the Molecular Basis for Metal Ion Effects on Plasminogen Activator Inhibitor-1 (PAI-1). [Doctoral Dissertation]. University of Tennessee – Knoxville; 2016. Available from: https://trace.tennessee.edu/utk_graddiss/3896

University of Tennessee – Knoxville
10.
Yue, Yufei.
Molecular Dynamics Simulations of Enzymes with Quantum Mechanical/Molecular Mechanical Potentials.
Degree: 2016, University of Tennessee – Knoxville
URL: https://trace.tennessee.edu/utk_graddiss/3984
► S-adenosyl methionine (SAM) dependent methylation process is universally found in all branches of life. It has important implications in mammalian pathogenesis and plant metabolism. The…
(more)
▼ S-adenosyl methionine (SAM) dependent methylation process is universally found in all branches of life. It has important implications in mammalian pathogenesis and plant metabolism. The methyl transfer is normally catalyzed by SAM-dependent methyltransferases(MTases). Two MTases are studied in this dissertation: the 1,7-dimethylxanthine methyltransferase (DXMT) which involve in plant caffeine biosynthesis, and the protein arginine methyltransferase 5(PRMT5) that participates in eukaryotic posttranslational modification. The late phase of caffeine biosynthesis starts from the substrate xanthosine and ends with the product caffeine, with theobromine as an intermediate product. DXMT is a key enzyme in this process and catalyzes two methylation steps: 1)methylation of 7-methylxanthine to form theobromine; 2)methylation of theobromine to form caffeine. The catalytic mechanism and product promiscuity of DXMT is intriguing. In Chapter 1, the quantum mechanical/molecular mechanical (QM/MM) molecular dynamics (MD) and free energy simulations were performed to explain the dual catalytic roles of DXMT. Simulation results show that a histidine residue may act as a general base catalyst during methylations. PRMTs can work as modifiers for histones and methylate the substrate arginine, thus interfering with histone code orchestration. The product specificity of PRMTs refers to their ability to produce either symmetric di-methylarginine(SDMA), asymmetric di-methylarginine(ADMA) or mono-methylarginine(MMA). Understanding the product specificity of PRMTs is important since different methylations may cause distinctive, even inverse biological consequences. PRMT5 produces SDMA, as compared to PRMT1 and PRMT3 that produce ADMA. In Chapter 2, simulations of PRMT5 have drawn a theoretical insight into the catalytic difference between SDMA and ADMA. Neddylation is a type of eukaryotic Ubiquitin-like (UBL) protein modification that is essential in cell division and development. Unlike ubiquitin and other small ubiquitin-like modifiers which target variety of protein substrates, the UBL NEDD8 is highly selective on modifying cullin proteins and contributes to 10% ~20% of all cellular ubiquitination and ubiquitination-like modification. In Chapter 3, the crystal structure of a trapped E3-E2 ̴ NEDD8-CUL1 intermediate was used for modeling, and simulations were applied to investigate the catalytic mechanism of NEDD8 transfer from E2 to the substrate. Some important insights were observed that may be used to understand the functional properties of the enzyme.
Subjects/Keywords: Biochemistry; Biophysics; and Structural Biology
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❌
APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
Yue, Y. (2016). Molecular Dynamics Simulations of Enzymes with Quantum Mechanical/Molecular Mechanical Potentials. (Doctoral Dissertation). University of Tennessee – Knoxville. Retrieved from https://trace.tennessee.edu/utk_graddiss/3984
Chicago Manual of Style (16th Edition):
Yue, Yufei. “Molecular Dynamics Simulations of Enzymes with Quantum Mechanical/Molecular Mechanical Potentials.” 2016. Doctoral Dissertation, University of Tennessee – Knoxville. Accessed March 04, 2021.
https://trace.tennessee.edu/utk_graddiss/3984.
MLA Handbook (7th Edition):
Yue, Yufei. “Molecular Dynamics Simulations of Enzymes with Quantum Mechanical/Molecular Mechanical Potentials.” 2016. Web. 04 Mar 2021.
Vancouver:
Yue Y. Molecular Dynamics Simulations of Enzymes with Quantum Mechanical/Molecular Mechanical Potentials. [Internet] [Doctoral dissertation]. University of Tennessee – Knoxville; 2016. [cited 2021 Mar 04].
Available from: https://trace.tennessee.edu/utk_graddiss/3984.
Council of Science Editors:
Yue Y. Molecular Dynamics Simulations of Enzymes with Quantum Mechanical/Molecular Mechanical Potentials. [Doctoral Dissertation]. University of Tennessee – Knoxville; 2016. Available from: https://trace.tennessee.edu/utk_graddiss/3984

Lehigh University
11.
Haaga, Jason Leigh.
Study of the Effects of Anisotropy in Self-Assembling Systems.
Degree: PhD, Physics, 2017, Lehigh University
URL: https://preserve.lehigh.edu/etd/4283
► There are many things in the world that are not spheres. As a result, isotropic interaction potentials can only serve as a crude approximation to…
(more)
▼ There are many things in the world that are not spheres. As a result, isotropic interaction potentials can only serve as a crude approximation to complex molecules such as proteins. In order to better understand the often harmful self-assembly phenomena that proteins can undergo, study of how anisotropic features alter collective behavior is required. Beyond the biological, these lessons can also guide rational design of new materials in the area of colloidal science. This dissertation examines the role of specific anisotropic features in coarse-grained representations of three proteins that undergo self-assembly processes. The first, amelogenin, is the primary protein involved in the formation of dental enamel; chapter 2 will explore the effect of the charged hydrophilic tail on the phase diagram of this otherwise hydrophobic protein. In chapter 3, the role of hinge angle between binding sites will be assessed in a simplified model of human antibodies; this angle has dramatic effect on aggregate morphology. Lastly, in chapter 4, the nucleation and growth mechanisms of polyglutamine tracts of different lengths will be studied, relevant to the formation of a class of neurodegenerative diseases, including Huntington's Disease. This process shows a strong dependence on repeat length, and at shorter lengths, variation due to concentration.
Advisors/Committee Members: Gunton, James D..
Subjects/Keywords: Biochemistry; Biophysics; and Structural Biology
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APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
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APA (6th Edition):
Haaga, J. L. (2017). Study of the Effects of Anisotropy in Self-Assembling Systems. (Doctoral Dissertation). Lehigh University. Retrieved from https://preserve.lehigh.edu/etd/4283
Chicago Manual of Style (16th Edition):
Haaga, Jason Leigh. “Study of the Effects of Anisotropy in Self-Assembling Systems.” 2017. Doctoral Dissertation, Lehigh University. Accessed March 04, 2021.
https://preserve.lehigh.edu/etd/4283.
MLA Handbook (7th Edition):
Haaga, Jason Leigh. “Study of the Effects of Anisotropy in Self-Assembling Systems.” 2017. Web. 04 Mar 2021.
Vancouver:
Haaga JL. Study of the Effects of Anisotropy in Self-Assembling Systems. [Internet] [Doctoral dissertation]. Lehigh University; 2017. [cited 2021 Mar 04].
Available from: https://preserve.lehigh.edu/etd/4283.
Council of Science Editors:
Haaga JL. Study of the Effects of Anisotropy in Self-Assembling Systems. [Doctoral Dissertation]. Lehigh University; 2017. Available from: https://preserve.lehigh.edu/etd/4283

University of Louisville
12.
Clark, Jennifer.
Enzyme kinetics : 6-phosphofructo-2-kinase/2,6-bisphosphatase.
Degree: MS, 2014, University of Louisville
URL: 10.18297/etd/1720
;
https://ir.library.louisville.edu/etd/1720
► Altered energy metabolism is an established hallmark of cancer cells. Fructose-2,6-bisphosphate is an allosteric activator of glycolysis and its concentration in a cell is…
(more)
▼ Altered energy metabolism is an established hallmark of cancer cells. Fructose-2,6-bisphosphate is an allosteric activator of glycolysis and its concentration in a cell is dictated by the 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatases (PFKFB1-4) family of bifunctional enzymes. The PFKFB family of enzymes are characterized by their different kinase: bisphosphatase activities and were originally determined to be tissue-specific. More recent data however suggest that multiple isoforms are co-expressed in both normal and neoplastic tissues. PFKFB4 is highly expressed in human cancer, strongly induced by hypoxia, and required for the survival of cancer cells. However, it remains unclear whether the kinase or phosphatase activity of human PFKFB4 dominates. In this study, we developed a method of synthesizing and purifying human recombinant PFKFB4 and, using this purified protein, found that PFKFB4 has a kinase to phosphatase ratio of 4.1:1. We found that a novel small molecule inhibitor of PFKFB4 (5MPN) inhibited the kinase activity of PFKFB4 and further that the known PFKFB3 inhibitors (PFK15 and PFK158) inhibited PFKFB4 less efficiently than 5MPN. We conclude that PFKFB4 functions mainly as a kinase and that, given the co-expression of PFKFB3 and PFKFB4, targeting both PFKFB3 and PFKFB4 isoforms using PFK158 and 5MPN may be a useful treatment option in cancer.
Advisors/Committee Members: Telang, Sucheta, Ellis, Steven, Clark, Barbara J..
Subjects/Keywords: Biochemistry; Biophysics; and Structural Biology
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APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
Clark, J. (2014). Enzyme kinetics : 6-phosphofructo-2-kinase/2,6-bisphosphatase. (Masters Thesis). University of Louisville. Retrieved from 10.18297/etd/1720 ; https://ir.library.louisville.edu/etd/1720
Chicago Manual of Style (16th Edition):
Clark, Jennifer. “Enzyme kinetics : 6-phosphofructo-2-kinase/2,6-bisphosphatase.” 2014. Masters Thesis, University of Louisville. Accessed March 04, 2021.
10.18297/etd/1720 ; https://ir.library.louisville.edu/etd/1720.
MLA Handbook (7th Edition):
Clark, Jennifer. “Enzyme kinetics : 6-phosphofructo-2-kinase/2,6-bisphosphatase.” 2014. Web. 04 Mar 2021.
Vancouver:
Clark J. Enzyme kinetics : 6-phosphofructo-2-kinase/2,6-bisphosphatase. [Internet] [Masters thesis]. University of Louisville; 2014. [cited 2021 Mar 04].
Available from: 10.18297/etd/1720 ; https://ir.library.louisville.edu/etd/1720.
Council of Science Editors:
Clark J. Enzyme kinetics : 6-phosphofructo-2-kinase/2,6-bisphosphatase. [Masters Thesis]. University of Louisville; 2014. Available from: 10.18297/etd/1720 ; https://ir.library.louisville.edu/etd/1720
13.
Liu, Dan.
Mitsunobu Reactions of Bulky Phenols and Lignin Model Dimer Synthesis & Decomposition.
Degree: MS, Chemistry and Biochemistry, 2015, South Dakota State University
URL: https://openprairie.sdstate.edu/etd/1808
► This thesis is divided into two parts: investigation of the Mitsunobu reaction of bulky phenols and aliphatic alcohols to improve yield, and studies of…
(more)
▼ This thesis is divided into two parts: investigation of the Mitsunobu reaction of bulky phenols and aliphatic alcohols to improve yield, and studies of lignin dimer model compound synthesis and catalysts of lignin decomposition reaction for optimizing degradation reaction conditions to increase product yields of value-added chemicals. In the first part of this thesis, Mitsunobu reaction of sterically hindered phenols and primary alcohols was investigated in detail to reveal the reason for the low yield of product (30-40%) and a solution to the problem was proposed and tested. In Mitsunobu alkyl aryl etherification reactions, alkylation of hydrazinedicarboxylate- a Mitsunobu by-product could be a side reaction. For essentially all Mitsunobu reactions in the literature, this side reaction is not a notable problem and good yields can be obtained with a wide range of solvents used. However, for the reactions of sterically hindered phenols and primary alcohols, this side reaction can significantly decrease the product yields. To suppress the side reaction and improve the product yields, solvent effect was studied and it was found that the yields are improved by using a weaker solvent, such as diethyl ether, instead of THF. In the second part of this thesis, synthesis of a lignin model compound (a dimer) and xiii study of its hydrothermal decomposition were conducted as an effort to explore effective methods to convert lignin, the largest renewable source of aromatics, into valuable chemicals. Since -O-4 bond is the predominant linkage in lignin, selective cleavage of this bond is sufficient to breakdown lignin into monomers and low oligomers which can be used directly for synthesis of certain materials such as adhesive resins. Lignin model compound with -O-4 bond has been synthesized through a four-step reaction scheme and the dimer decomposition using various catalysts has been studied. It is found that repolymerization (or condensation) happens at a relatively low temperature (120 °C) with or without a catalyst. This work points out the need to develop methods to suppress condensation reactions.
Advisors/Committee Members: Cheng Zhang.
Subjects/Keywords: Biochemistry; Biophysics; and Structural Biology
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APA ·
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MLA ·
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CSE |
Export
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APA (6th Edition):
Liu, D. (2015). Mitsunobu Reactions of Bulky Phenols and Lignin Model Dimer Synthesis & Decomposition. (Masters Thesis). South Dakota State University. Retrieved from https://openprairie.sdstate.edu/etd/1808
Chicago Manual of Style (16th Edition):
Liu, Dan. “Mitsunobu Reactions of Bulky Phenols and Lignin Model Dimer Synthesis & Decomposition.” 2015. Masters Thesis, South Dakota State University. Accessed March 04, 2021.
https://openprairie.sdstate.edu/etd/1808.
MLA Handbook (7th Edition):
Liu, Dan. “Mitsunobu Reactions of Bulky Phenols and Lignin Model Dimer Synthesis & Decomposition.” 2015. Web. 04 Mar 2021.
Vancouver:
Liu D. Mitsunobu Reactions of Bulky Phenols and Lignin Model Dimer Synthesis & Decomposition. [Internet] [Masters thesis]. South Dakota State University; 2015. [cited 2021 Mar 04].
Available from: https://openprairie.sdstate.edu/etd/1808.
Council of Science Editors:
Liu D. Mitsunobu Reactions of Bulky Phenols and Lignin Model Dimer Synthesis & Decomposition. [Masters Thesis]. South Dakota State University; 2015. Available from: https://openprairie.sdstate.edu/etd/1808

University of Oxford
14.
Alderson, Thomas Reid.
Protein folding investigated by NMR spectroscopy.
Degree: PhD, 2019, University of Oxford
URL: http://ora.ox.ac.uk/objects/uuid:d09c75a9-c8d5-422b-8a69-79a3cd3c252f
;
https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.813641
► Proteins are biological molecules that perform diverse cellular roles, including metabolizing energy sources to regulating the acidity of blood. The collective actions of proteins and…
(more)
▼ Proteins are biological molecules that perform diverse cellular roles, including metabolizing energy sources to regulating the acidity of blood. The collective actions of proteins and their interactions with nucleic acids at the molecular level enable life at the macroscopic level. If protein folding is delayed or has faltered, unwanted associations between exposed hydrophobic amino acids can lead to cytotoxic protein aggregation, which is implicated in diseases such as Parkinson’s, Alzheimer’s, and type II diabetes. However, cells have armed themselves with an evolutionarily conserved defense mechanism to combat protein misfolding: a class of proteins known as molecular chaperones that can recognize misfolded, aggregation-prone proteins and either correctly refold them or target them for degradation and recycling. The focus of this thesis is to understand aspects of protein folding and the activity of molecular chaperones at the atomic level. Nuclear magnetic resonance (NMR) spectroscopy methods have been applied to characterize the molecular chaperone HSP27. My results reveal that HSP27 locally unfolds upon monomerization and becomes highly active. Further, I characterize a single amino acid variant of HSP27 implicated in the onset of Charcot-Marie-Tooth disease, a commonly inherited motor neuropathy, and find to be devoid of chaperone activity, while also forming oligomers that are significantly larger than the wild-type protein. Rapid pressure-jump NMR experiments are developed and deployed to monitor protein folding with millisecond resolution via the protection from solvent exchange. These data provide insight into the folding mechanism of a pressure-sensitized variant of ubiquitin. The extent of cis-proline formation is characterized in unfolded proteins with NMR, and the degree of polypeptide collapse in low denaturant conditions is probed by translational diffusion. Finally, the accuracy and precision of Carr-Purcell-Meiboom-Gill relaxation dispersion, a powerful NMR method to investigate transiently populated folding intermediates, is analyzed in the presence of limited input data.
Subjects/Keywords: Structural Biology; Biochemistry; Biophysics
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APA ·
Chicago ·
MLA ·
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CSE |
Export
to Zotero / EndNote / Reference
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APA (6th Edition):
Alderson, T. R. (2019). Protein folding investigated by NMR spectroscopy. (Doctoral Dissertation). University of Oxford. Retrieved from http://ora.ox.ac.uk/objects/uuid:d09c75a9-c8d5-422b-8a69-79a3cd3c252f ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.813641
Chicago Manual of Style (16th Edition):
Alderson, Thomas Reid. “Protein folding investigated by NMR spectroscopy.” 2019. Doctoral Dissertation, University of Oxford. Accessed March 04, 2021.
http://ora.ox.ac.uk/objects/uuid:d09c75a9-c8d5-422b-8a69-79a3cd3c252f ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.813641.
MLA Handbook (7th Edition):
Alderson, Thomas Reid. “Protein folding investigated by NMR spectroscopy.” 2019. Web. 04 Mar 2021.
Vancouver:
Alderson TR. Protein folding investigated by NMR spectroscopy. [Internet] [Doctoral dissertation]. University of Oxford; 2019. [cited 2021 Mar 04].
Available from: http://ora.ox.ac.uk/objects/uuid:d09c75a9-c8d5-422b-8a69-79a3cd3c252f ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.813641.
Council of Science Editors:
Alderson TR. Protein folding investigated by NMR spectroscopy. [Doctoral Dissertation]. University of Oxford; 2019. Available from: http://ora.ox.ac.uk/objects/uuid:d09c75a9-c8d5-422b-8a69-79a3cd3c252f ; https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.813641

University of Colorado
15.
Alzahrani, Abeer Ahmed.
Copper-Catalyzed Azide Alkyne Cycloaddition Polymer Networks.
Degree: PhD, Chemistry & Biochemistry, 2014, University of Colorado
URL: https://scholar.colorado.edu/chem_gradetds/1
► The click reaction concept, introduced in 2001, has since spurred the rapid development and reexamination of efficient, high yield reactions which proceed rapidly under…
(more)
▼ The click reaction concept, introduced in 2001, has since spurred the rapid development and reexamination of efficient, high yield reactions which proceed rapidly under mild conditions. Prior to the discovery of facile copper catalysis in 2002, the thermally activated azidealkyne or Huisgen cycloaddition reaction was largely ignored following its discovery in large part due to its slow kinetics, requirement for elevated temperature and limited selectivity. Now, arguably, the most prolific and capable of the click reactions, the copper-catalyzed azide alkyne cycloaddition (CuAAC) reaction is extremely efficient and affords exquisite control of the reaction. The orthogonally and chemoselectivity of this reaction enable its wide utility across varied scientific fields. Despite numerous inherent advantages and widespread use for small molecule synthesis and solution-based polymer chemistry, it has only recently and rarely been utilized to form polymer networks. This work focuses on the synthesis, mechanisms, and unique attributes of the CuAAC reaction for the fabrication of functional polymer networks.
The photo-reduction of a series of copper(II)/amine complexes via ligand metal charge transfer was examined to determine their relative efficiency and selectivity in catalyzing the CuAAC reaction. The aliphatic amine ligands were used as an electron transfer species to reduce Cu(II) upon irradiation with 365 nm light while also functioning as an accelerating agent and as protecting ligands for the Cu(I) that was formed. Among the aliphatic amines studied, tertiary amines such as triethylamine (TEA), tetramethyldiamine (TMDA), N,N,N’,N”,N”-pentamethyldiethylenetriamine (PMDTA), and hexamethylenetetramine (HMTETA) were found to be the most effective. The reaction kinetics were accelerated by increasing the PMDETA : Cu(II) ratio with a ratio of ligand to Cu(II) of 4:1 yielding the maximum conversion in the shortest time. The sequential and orthogonal nature of the photo-CuAAC reaction and a chain-growth acrylate homopolymerization were demonstrated and used to form branched polymer structures.
A bulk, organic soluble initiation system consisting of a Cu(II) salt and a primary amine was also examined in both model reactions and in bulk polymerizations. The system was shown to be highly efficient, leading to nearly complete CuAAC polymerization at ambient temperature. Increasing the ratio of amine to copper from 1 to 4 increases the CuAAC reaction rate significantly from 4 mM/min for 1:1 ratio of Cu(II):hexyalmine to 14mM/min for 1:4 ratio. The concentration dependence of the amine on the reaction rate enables the polymerization rate to be controlled simply by manipulating the hexylamine concentration.
Sequential thiol–acrylate and photo-CuAAC click reactions were utilized to form twostage reactive polymer networks capable of generating wrinkles in a facile manner. The click thiol- Michael addition reaction was utilized to form a cross-linked polymer with residual, reactive alkyne sites that remained…
Advisors/Committee Members: Christopher N. Bowman, Jeffrey W. Stansbury.
Subjects/Keywords: Biochemistry; Biochemistry, Biophysics, and Structural Biology; Chemistry
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❌
APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
Alzahrani, A. A. (2014). Copper-Catalyzed Azide Alkyne Cycloaddition Polymer Networks. (Doctoral Dissertation). University of Colorado. Retrieved from https://scholar.colorado.edu/chem_gradetds/1
Chicago Manual of Style (16th Edition):
Alzahrani, Abeer Ahmed. “Copper-Catalyzed Azide Alkyne Cycloaddition Polymer Networks.” 2014. Doctoral Dissertation, University of Colorado. Accessed March 04, 2021.
https://scholar.colorado.edu/chem_gradetds/1.
MLA Handbook (7th Edition):
Alzahrani, Abeer Ahmed. “Copper-Catalyzed Azide Alkyne Cycloaddition Polymer Networks.” 2014. Web. 04 Mar 2021.
Vancouver:
Alzahrani AA. Copper-Catalyzed Azide Alkyne Cycloaddition Polymer Networks. [Internet] [Doctoral dissertation]. University of Colorado; 2014. [cited 2021 Mar 04].
Available from: https://scholar.colorado.edu/chem_gradetds/1.
Council of Science Editors:
Alzahrani AA. Copper-Catalyzed Azide Alkyne Cycloaddition Polymer Networks. [Doctoral Dissertation]. University of Colorado; 2014. Available from: https://scholar.colorado.edu/chem_gradetds/1
16.
Kudire, Anay Kumar.
Synthesis of Functionalized Carboxylate Deposition Materials for DSSC and New Chromatography Stationary Phases.
Degree: MS, Chemistry and Biochemistry, 2015, South Dakota State University
URL: https://openprairie.sdstate.edu/etd/1821
► Hybrid materials take advantage of the unique properties of two single-phase materials (e.g., organic and inorganic) by bonding these materials together, producing a two-phase…
(more)
▼ Hybrid materials take advantage of the unique properties of two single-phase materials (e.g., organic and inorganic) by bonding these materials together, producing a two-phase material with properties that cannot be achieved by a single-phase material. In this study, a set of hybrid materials were synthesized for deposition via Functionalized Carboxylate Deposition (FCD), a novel gas-phase deposition technique developed in our laboratory, for use in dye sensitized solar cell (DSSC) and chromatography applications. FCD-compatible ruthenium dye complexes were synthesized based on conventional DSSC dyes (N3 and N719). These dyes were modified with cyano-acrylic groups (CAA) to form N3-CAA and N719-CAA dyes. Synthesis was accomplished by initially oxidizing a bipyridine dimethanol ligand and using Knoevenagel condensation to introduce CAA functionality. These dyes produced efficiencies of 1.4-1.5%. Organic bonded phases were similarly synthesized for application as chromatographic stationary phases. 2-cyano 3-phenyl acrylic acid (CPA), 2-cyano 3-pentaflorophynyl acrylic acids (CFA), 2-cyano hexenic acid (2-CHeA), 2-cyano hexanoic acid (2-CHaA) hybrid molecules were synthesized via Knoevenagel condensation of their corresponding aldehyde precursors. The resulting compounds were deposited on silica solid support materials and the chromatographic behavior of methyl violet using the novel stationary phases was evaluated.
Advisors/Committee Members: Brian A. Logue.
Subjects/Keywords: Biochemistry; Biochemistry, Biophysics, and Structural Biology; Chemistry
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MLA ·
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CSE |
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APA (6th Edition):
Kudire, A. K. (2015). Synthesis of Functionalized Carboxylate Deposition Materials for DSSC and New Chromatography Stationary Phases. (Masters Thesis). South Dakota State University. Retrieved from https://openprairie.sdstate.edu/etd/1821
Chicago Manual of Style (16th Edition):
Kudire, Anay Kumar. “Synthesis of Functionalized Carboxylate Deposition Materials for DSSC and New Chromatography Stationary Phases.” 2015. Masters Thesis, South Dakota State University. Accessed March 04, 2021.
https://openprairie.sdstate.edu/etd/1821.
MLA Handbook (7th Edition):
Kudire, Anay Kumar. “Synthesis of Functionalized Carboxylate Deposition Materials for DSSC and New Chromatography Stationary Phases.” 2015. Web. 04 Mar 2021.
Vancouver:
Kudire AK. Synthesis of Functionalized Carboxylate Deposition Materials for DSSC and New Chromatography Stationary Phases. [Internet] [Masters thesis]. South Dakota State University; 2015. [cited 2021 Mar 04].
Available from: https://openprairie.sdstate.edu/etd/1821.
Council of Science Editors:
Kudire AK. Synthesis of Functionalized Carboxylate Deposition Materials for DSSC and New Chromatography Stationary Phases. [Masters Thesis]. South Dakota State University; 2015. Available from: https://openprairie.sdstate.edu/etd/1821
17.
Huang, Lu.
CBL and CBL-B Dictate CSF-1R Endocytic Traffic and Signaling in Macrophages.
Degree: PhD, Chemistry and Biochemistry, 2017, South Dakota State University
URL: https://openprairie.sdstate.edu/etd/1198
► Macrophage colony stimulating factor receptor (CSF-1R or MCSFR) is a receptor tyrosine kinase essential for the growth and function of macrophages. Understanding the mechanisms…
(more)
▼ Macrophage colony stimulating factor receptor (CSF-1R or MCSFR) is a receptor tyrosine kinase essential for the growth and function of macrophages. Understanding the mechanisms that regulate CSF-1R activation and deactivation will provide insights to clinical treatment of macrophage related diseases including chronic inflammation and cancer. Previously, our laboratory showed that CSF-1R undergoes a novel membrane trafficking route that involves macropinocytosis to deactivate CSF-1R signaling. This thesis makes the discovery that the ubiquitin ligases Cbl and Cbl-b cooperate to regulate CSF-1R endocytosis and traffic to macropinosome in macrophages. Macrophages were derived from mice knocked out for Cbl, Cbl-b or the double knock out (DKO). DKO macrophages hyperproliferated, matching the severe myeloproliferative disorder observed in DKO mice. The CSF-1R and associated proteins were not ubiquitinated in DKO macrophages, unlike single knockouts and wild-type cells suggesting redundant functions of Cbl and Cbl-b. Mapping of the CSF-1R traffic demonstrated that CSF-1R internalization was slower in DKO cells, resulting in prolonged CSF-1R signaling at the plasma membrane and prolonged Akt signaling. Interestingly, CSF-1R transport to the lumen of macropinosome was defective in DKO cells, suggesting that altered membrane transport is responsible for the cellular phenotype. Tyrosine phosphorylation was drastically decreased and ERK signaling was lower in DKO macrophages, possibly x resulting from defective CSF-1R signaling on endosome and macropinosome. One of the key ESCRT proteins, HRS, did not associate with the CSF-1R in DKO macrophages indicating Cbl and Cbl-b are required for ESCRT-mediated transport into the lumen of macropinosome. Surprisingly, the CSF-1R was still degraded in DKO cells by an unknown mechanism. RNA sequencing analysis showed that Cbl and Cbl-b work together to regulate approximately 1,300 genes, while Cbl appears to regulate a unique set of approximately 250 genes in macrophages. In conclusion, Cbl and Cbl-b share partially redundant functions regulating CSF-1R signaling, endocytic traffic and cell growth.
Advisors/Committee Members: Adam Hoppe.
Subjects/Keywords: Biochemistry; Biochemistry, Biophysics, and Structural Biology; Chemistry
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❌
APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
Huang, L. (2017). CBL and CBL-B Dictate CSF-1R Endocytic Traffic and Signaling in Macrophages. (Doctoral Dissertation). South Dakota State University. Retrieved from https://openprairie.sdstate.edu/etd/1198
Chicago Manual of Style (16th Edition):
Huang, Lu. “CBL and CBL-B Dictate CSF-1R Endocytic Traffic and Signaling in Macrophages.” 2017. Doctoral Dissertation, South Dakota State University. Accessed March 04, 2021.
https://openprairie.sdstate.edu/etd/1198.
MLA Handbook (7th Edition):
Huang, Lu. “CBL and CBL-B Dictate CSF-1R Endocytic Traffic and Signaling in Macrophages.” 2017. Web. 04 Mar 2021.
Vancouver:
Huang L. CBL and CBL-B Dictate CSF-1R Endocytic Traffic and Signaling in Macrophages. [Internet] [Doctoral dissertation]. South Dakota State University; 2017. [cited 2021 Mar 04].
Available from: https://openprairie.sdstate.edu/etd/1198.
Council of Science Editors:
Huang L. CBL and CBL-B Dictate CSF-1R Endocytic Traffic and Signaling in Macrophages. [Doctoral Dissertation]. South Dakota State University; 2017. Available from: https://openprairie.sdstate.edu/etd/1198

University of Tennessee – Knoxville
18.
Raval, Sherin R.
A step towards understanding of the molecular basis of ligand promiscuity in the aminoglycoside modifying enzymes.
Degree: MS, Biochemistry and Cellular and Molecular Biology, 2014, University of Tennessee – Knoxville
URL: https://trace.tennessee.edu/utk_gradthes/3173
► Aminoglycosides have proven very useful in the treatment of infections; lately their effectiveness has been greatly reduced due to increasing resistance. Among many known…
(more)
▼ Aminoglycosides have proven very useful in the treatment of infections; lately their effectiveness has been greatly reduced due to increasing resistance. Among many known mechanisms of resistance to aminoglycosides, enzymatic modification is the most prevailing. More than 14 aminoglycoside -
N3-acetyltransferases- a class of aminoglycoside modifying enzymes, are known today. This study focuses on a pair of acetyl transferases: The aminoglycoside-
N3- acetyltransferase IIIb (AAC-IIIb) and the aminoglycoside-
N3- acetyltransferase IIa (AAC-IIa). AAC-IIa and AAC-IIIb are very similar in their amino acid sequence and structure – yet they have a strong difference in their substrate selectivity, kinetic and thermodynamic properties. This work represents a comparative study of these two enzymes in an effort to determine thermodynamic basis of the differential substrate profiles of AAC-IIa to AAC-IIIb.
Advisors/Committee Members: Engin H. Serpersu, Gladys Alexandre, Hong Guo.
Subjects/Keywords: Biochemistry, Biophysics, and Structural Biology; Molecular Biology
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APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
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APA (6th Edition):
Raval, S. R. (2014). A step towards understanding of the molecular basis of ligand promiscuity in the aminoglycoside modifying enzymes. (Thesis). University of Tennessee – Knoxville. Retrieved from https://trace.tennessee.edu/utk_gradthes/3173
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):
Raval, Sherin R. “A step towards understanding of the molecular basis of ligand promiscuity in the aminoglycoside modifying enzymes.” 2014. Thesis, University of Tennessee – Knoxville. Accessed March 04, 2021.
https://trace.tennessee.edu/utk_gradthes/3173.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Raval, Sherin R. “A step towards understanding of the molecular basis of ligand promiscuity in the aminoglycoside modifying enzymes.” 2014. Web. 04 Mar 2021.
Vancouver:
Raval SR. A step towards understanding of the molecular basis of ligand promiscuity in the aminoglycoside modifying enzymes. [Internet] [Thesis]. University of Tennessee – Knoxville; 2014. [cited 2021 Mar 04].
Available from: https://trace.tennessee.edu/utk_gradthes/3173.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Raval SR. A step towards understanding of the molecular basis of ligand promiscuity in the aminoglycoside modifying enzymes. [Thesis]. University of Tennessee – Knoxville; 2014. Available from: https://trace.tennessee.edu/utk_gradthes/3173
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
19.
Millage, Megan R.
Rab39 and Klp98A are Required for Furrow Formation During Early Drosophila Embryogenesis.
Degree: M. S., Biological Sciences, 2020, U of Denver
URL: https://digitalcommons.du.edu/etd/1812
► The formation of a plasma membrane furrow is an essential process during development. Furrow formation is necessary for successful cell division and cytokinesis in…
(more)
▼ The formation of a plasma membrane furrow is an essential process during development. Furrow formation is necessary for successful cell division and cytokinesis in addition to the ability to create multicellular tissues. Here, I will explore the role of the Golgi-associated Rab protein Rab39 in furrow formation during early Drosophila embryogenesis. Rab39 is one of eight Rab proteins that has been shown to localize to discrete puncta by live imaging in early Drosophila embryos, but its function and pathway have not been well characterized. In this thesis will I show that Rab39 forms dynamic, tubular structures that colocalize with trans-Golgi markers and the knockdown of Rab39 using RNA interference causes defects in furrow length and nuclear division during syncytial cycles 10-13. Klp98A, a kinesin 3 family motor protein, produces similar abnormalities when disrupted and colocalizes with Rab39. In the absence of Rab39, Klp98A strongly relocalizes to large nuclear fragments outside of the nuclear envelope that have arisen from the defects in furrow formation. Additionally, Rab39 and Klp98A dynamics are dependent on microtubule networks. Together, these proteins could represent a novel pathway that mediate membrane trafficking from the Golgi to the plasma membrane to aid in furrow formation.
Advisors/Committee Members: J. Todd Blankenship, Dinah Loerke, Cedric Asensio.
Subjects/Keywords: Biochemistry, Biophysics, and Structural Biology; Molecular Biology
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❌
APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
Millage, M. R. (2020). Rab39 and Klp98A are Required for Furrow Formation During Early Drosophila Embryogenesis. (Thesis). U of Denver. Retrieved from https://digitalcommons.du.edu/etd/1812
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):
Millage, Megan R. “Rab39 and Klp98A are Required for Furrow Formation During Early Drosophila Embryogenesis.” 2020. Thesis, U of Denver. Accessed March 04, 2021.
https://digitalcommons.du.edu/etd/1812.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Millage, Megan R. “Rab39 and Klp98A are Required for Furrow Formation During Early Drosophila Embryogenesis.” 2020. Web. 04 Mar 2021.
Vancouver:
Millage MR. Rab39 and Klp98A are Required for Furrow Formation During Early Drosophila Embryogenesis. [Internet] [Thesis]. U of Denver; 2020. [cited 2021 Mar 04].
Available from: https://digitalcommons.du.edu/etd/1812.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Millage MR. Rab39 and Klp98A are Required for Furrow Formation During Early Drosophila Embryogenesis. [Thesis]. U of Denver; 2020. Available from: https://digitalcommons.du.edu/etd/1812
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Florida International University
20.
cao, nan.
Structure and Mechanism of Mycobacterial Topoisomerase I.
Degree: PhD, Biochemistry, 2018, Florida International University
URL: https://digitalcommons.fiu.edu/etd/3747
;
10.25148/etd.FIDC006890
;
FIDC006890
► The enzyme DNA topoisomerase I is an essential enzyme that plays an important role in eukaryotic and prokaryotic cellular processes such as DNA replication,…
(more)
▼ The enzyme DNA topoisomerase I is an essential enzyme that plays an important role in eukaryotic and prokaryotic cellular processes such as DNA replication, transcription, recombination and repair.
Mycobacterium tuberculosistopoisomerase I (MtTOP1) is a validated drug target for antituberculosis treatment. Mycobacterial topoisomerase I regulates the topological constraints in chromosomes and helps in maintaining the growth of mycobacteria. The N- terminal domain (NTD) of mycobacterial topoisomerase I contains conserved catalytic domains that along with the active site Tyrosine are involved in cleaving and rejoining a single strand of DNA. Magnesium is required in DNA cleavage activity of type IA topoisomerases. The C-terminal domain (CTD) of mycobacterial topoisomerase I is divided into four subdomains (D5-D8) and a positively charged tail. Each subdomain has a GxxGPY sequence motif. The DNA binding, relaxation, cleavage, religation, catenation and decatenation ability of each subdomains of CTD were studied. The present study shows that each subdomain has its own characteristics. Subdomain D8 and D7 are responsible for maintaining the relaxation activity of mycobacterial topoisomerase I. Subdomain D5 is essential to maintain the DNA cleavage, religation, catenation and decatenation activity. A new crystal structure of MtTOP1-704t (amino acids A2-T704 containing NTD+D5 domains) was obtained. Structures with ssDNA substrate bound to the active site (Y342) in the presence and absence of Mg2+ were also investigated. Significant enzyme conformational changes upon DNA binding place the catalytic tyrosine in a pre-transition position for cleavage of a specific phosphodiester linkage to form a covalent intermediate. Meanwhile, the enzyme/DNA complex with Mg2+ bound at active site may present the post- transition state for religation in the enzyme’s multiple-state DNA relaxation activity. The critical function of a strictly conserved glutamic acid in acid-base catalysis of the DNA cleavage step was also demonstrated by site-directed mutagenesis. The present work provides new functional insights into the more stringent requirement for DNA rejoining versus cleavage by type IA topoisomerase, and further establishes the potential for select interference of DNA rejoining via specific inhibitors.
Advisors/Committee Members: Yuk-Ching Tse-Dinh, Xiaotang Wang, Yuan Liu, Watson Lees, Kalai Mathee.
Subjects/Keywords: Biochemistry; Molecular Biology; Biophysics; Biochemistry; Molecular Biology; Other Biochemistry, Biophysics, and Structural Biology
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❌
APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
cao, n. (2018). Structure and Mechanism of Mycobacterial Topoisomerase I. (Doctoral Dissertation). Florida International University. Retrieved from https://digitalcommons.fiu.edu/etd/3747 ; 10.25148/etd.FIDC006890 ; FIDC006890
Chicago Manual of Style (16th Edition):
cao, nan. “Structure and Mechanism of Mycobacterial Topoisomerase I.” 2018. Doctoral Dissertation, Florida International University. Accessed March 04, 2021.
https://digitalcommons.fiu.edu/etd/3747 ; 10.25148/etd.FIDC006890 ; FIDC006890.
MLA Handbook (7th Edition):
cao, nan. “Structure and Mechanism of Mycobacterial Topoisomerase I.” 2018. Web. 04 Mar 2021.
Vancouver:
cao n. Structure and Mechanism of Mycobacterial Topoisomerase I. [Internet] [Doctoral dissertation]. Florida International University; 2018. [cited 2021 Mar 04].
Available from: https://digitalcommons.fiu.edu/etd/3747 ; 10.25148/etd.FIDC006890 ; FIDC006890.
Council of Science Editors:
cao n. Structure and Mechanism of Mycobacterial Topoisomerase I. [Doctoral Dissertation]. Florida International University; 2018. Available from: https://digitalcommons.fiu.edu/etd/3747 ; 10.25148/etd.FIDC006890 ; FIDC006890

University of California – Berkeley
21.
Chao, Luke H.
Structural Studies of Calcium/Calmodulin Depenedent Protein Kinase II Activation.
Degree: Molecular & Cell Biology, 2010, University of California – Berkeley
URL: http://www.escholarship.org/uc/item/9bt8t2n7
► Cell signaling utilizes the frequency of calcium stimuli to produce diverse physiological outcomes. From fertilization of oocytes and cardiac facilitation, to muscle contraction and action…
(more)
▼ Cell signaling utilizes the frequency of calcium stimuli to produce diverse physiological outcomes. From fertilization of oocytes and cardiac facilitation, to muscle contraction and action potential firing, periodic fluctuations in calcium levels play a key role in determining cell fate. During my thesis work, I studied a protein signaling complex that responds to the frequency of calcium stimuli: calcium/calmodulin dependent protein kinase II (CaMKII). The CaMKII holoenzyme is a dodecameric kinase assembly that activates and autophosphorylates in a manner dependent on the frequency of calcium stimuli. Its best-studied role is in the post-synaptic neuron, where it responds to calcium pulse frequencies to initiate changes important for Long Term Potentiation.My work investigated the activation mechanism of CaMKII, with the goal of understanding the enzyme's cooperative response to calcium-saturated calmodulin. This work elucidates the mechanism for CaMKII activation; by demonstrating that CaMKII is not a simple `coincidence detector' as previously postulated, by showing that CaMKII is cooperatively activated by calcium-saturated calmodulin, and by demonstrating that cooperative activation is mediated by the inter-subunit `capture' of regulatory segments. This work demonstrates that the cooperative response of CaMKII can by modulated by the length of the linker region where splice-form deletions and insertions occur. Similar modifications have been shown to shift the frequency-dependent activation of the enzyme, suggesting that our proposed mechanism enables the enzyme to tune its frequency response depending on developmental or tissue-specific expression. In addition, analysis of interactions exhibited by a peptide inhibitor of CaMKII revealed docking sites which other proteins, such as the NMDA-receptor, may utilize in order to localize and affect the frequency response of the CaMKII. Finally, I investigated the exchange of CaMKII subunits between holoenzyme complexes, and discuss the implications of such a process in prolonging the response to calcium stimuli beyond the lifespan of individual protein subunits.
Subjects/Keywords: Biophysics, General; Chemistry, Biochemistry; CaMKII; Structural Biology
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❌
APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
Chao, L. H. (2010). Structural Studies of Calcium/Calmodulin Depenedent Protein Kinase II Activation. (Thesis). University of California – Berkeley. Retrieved from http://www.escholarship.org/uc/item/9bt8t2n7
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):
Chao, Luke H. “Structural Studies of Calcium/Calmodulin Depenedent Protein Kinase II Activation.” 2010. Thesis, University of California – Berkeley. Accessed March 04, 2021.
http://www.escholarship.org/uc/item/9bt8t2n7.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Chao, Luke H. “Structural Studies of Calcium/Calmodulin Depenedent Protein Kinase II Activation.” 2010. Web. 04 Mar 2021.
Vancouver:
Chao LH. Structural Studies of Calcium/Calmodulin Depenedent Protein Kinase II Activation. [Internet] [Thesis]. University of California – Berkeley; 2010. [cited 2021 Mar 04].
Available from: http://www.escholarship.org/uc/item/9bt8t2n7.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Chao LH. Structural Studies of Calcium/Calmodulin Depenedent Protein Kinase II Activation. [Thesis]. University of California – Berkeley; 2010. Available from: http://www.escholarship.org/uc/item/9bt8t2n7
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Virginia Commonwealth University
22.
Van, Danielle.
Determining the mechanism of double-stranded RNA-induced cell death in ovarian cancer.
Degree: PhD, Biochemistry, 2011, Virginia Commonwealth University
URL: https://doi.org/10.25772/2WAV-E830
;
https://scholarscompass.vcu.edu/etd/265
► Ovarian cancer is the most lethal of all gynecological cancers. Current ovarian cancer drug regimens, including taxanes and platinum-based agents, are susceptible to chemoresistance necessitating…
(more)
▼ Ovarian cancer is the most lethal of all gynecological cancers. Current ovarian cancer drug regimens, including taxanes and platinum-based agents, are susceptible to chemoresistance necessitating the development of novel chemotherapeutics. Within tumors pathogen-derived ligands, such as dsRNA, can activate pattern recognition receptors (PRRs) that are capable of inducing apoptosis. In this dissertation we have found that in ovarian cancer cell lines (DOV-13, SKOV-3, CAOV-3, and OVCAR-3), dsRNA treatment alters cell survival. When treated with dsRNA, ovarian cancer cell lines and patient samples could be divided into two categories, responsive which undergo significant levels of apoptosis (CAOV-3 and OVCAR-3) or non-responsive which are unaffected (DOV-13 and SKOV-3). Following dsRNA treatment, dsRNA receptor expression levels increase in responsive cell lines and patient samples only. This suggests a potential role for dsRNA receptors as biomarkers to identify dsRNA-responsive patients. Detailed investigation of the mechanism by which cell lines succumb to or avoid dsRNA-induced cell death showed that in responsive cell lines, NF-kappaB, IFN-beta and caspase 3 activation occurred. Cell death was caspase and IFN-dependent. In non-responsive cell lines, increased c-IAP2 levels and RIP1 kinase ubiquitination occurred, as well as, an increase in basal level autophagy with dsRNA stimulation. However, individual blockade of these pathways did not restore dsRNA-induced apoptosis. In a non-responsive cell line, dsRNA enhanced the action of paclitaxel, carboplatin, and vorinostat through an as yet undetermined mechanism. In a responsive cell line, dsRNA produced a synergistic effect when combined with these drugs. These novel dual therapies, innate immune ligand plus cytotoxic agent, may find application in chemoresistant ovarian cancers.
Advisors/Committee Members: Jessica Bell.
Subjects/Keywords: Biochemistry, Biophysics, and Structural Biology; Life Sciences
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❌
APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
Van, D. (2011). Determining the mechanism of double-stranded RNA-induced cell death in ovarian cancer. (Doctoral Dissertation). Virginia Commonwealth University. Retrieved from https://doi.org/10.25772/2WAV-E830 ; https://scholarscompass.vcu.edu/etd/265
Chicago Manual of Style (16th Edition):
Van, Danielle. “Determining the mechanism of double-stranded RNA-induced cell death in ovarian cancer.” 2011. Doctoral Dissertation, Virginia Commonwealth University. Accessed March 04, 2021.
https://doi.org/10.25772/2WAV-E830 ; https://scholarscompass.vcu.edu/etd/265.
MLA Handbook (7th Edition):
Van, Danielle. “Determining the mechanism of double-stranded RNA-induced cell death in ovarian cancer.” 2011. Web. 04 Mar 2021.
Vancouver:
Van D. Determining the mechanism of double-stranded RNA-induced cell death in ovarian cancer. [Internet] [Doctoral dissertation]. Virginia Commonwealth University; 2011. [cited 2021 Mar 04].
Available from: https://doi.org/10.25772/2WAV-E830 ; https://scholarscompass.vcu.edu/etd/265.
Council of Science Editors:
Van D. Determining the mechanism of double-stranded RNA-induced cell death in ovarian cancer. [Doctoral Dissertation]. Virginia Commonwealth University; 2011. Available from: https://doi.org/10.25772/2WAV-E830 ; https://scholarscompass.vcu.edu/etd/265

Virginia Commonwealth University
23.
Bhardwaj, Reetika.
REGULATION OF YKL-40 IN STERILE INFLAMMATION AND ITS ROLE IN GLIOBLASTOMA IN VIVO.
Degree: PhD, Biochemistry, 2014, Virginia Commonwealth University
URL: https://doi.org/10.25772/4JHN-WG10
;
https://scholarscompass.vcu.edu/etd/629
► YKL-40 is a secreted glycoprotein, which is a shared biomarker of chronic inflammation and oncogenic transformation. Indeed, YKL-40 expression is up-regulated in many diseases…
(more)
▼ YKL-40 is a secreted glycoprotein, which is a shared biomarker of chronic inflammation and oncogenic transformation. Indeed, YKL-40 expression is up-regulated in many diseases including multiple sclerosis, Alzheimer disease, viral encephalitis, HIV-associated dementia, brain infarction, and traumatic brain injury. YKL-40 is also expressed by several solid tumors, such as osteosarcoma, ovarian carcinoma and glioblastoma multiforme (GBM). It promotes the migration and invasion of astrocytes as well as GBM cells. Serum YKL-40 levels have been shown to directly correlate with tumor grade and potentially tumor burden in GBM. In contrast to the numerous reports documenting elevated expression of YKL-40, relatively little is known about inflammatory mediators and specific molecular mechanisms that control its expression. For my PhD project, I decided to elucidate the mechanism regulating YKL-40 expression in inflammation and to understand how YKL-40 mediates the migration and invasion of GBM cells in vivo. As per my first project, I found that YKL-40 expression is up-regulated in many inflammatory models such as irritant induced inflammation (turpentine), EAE (experimental autoimmune encephalomyelitis), irradiation induced brain inflammation and in the HIV-TAT overexpression model. YKL-40 expression is also up regulated in oligodendroglioma patient samples. We show that YKL-40 expression is activated by major pro-inflammatory cytokines including IL-1, IL-6, and OSM in astrocytes, and that the activation of YKL-40 expression by cytokines depends on the STAT and NF-κB regulatory elements located within the YKL-40 promoter. Additionally, we discovered that STAT3 is a major regulator of YKL-40 expression in response to the IL-6 family cytokines, including OSM. Indeed, both depletion of STAT3 expression or overexpression of dominant-negative STAT3 abolishes activation of YKL-40 expression. In contrast, although IL-1 activates NF-κB (p65/p50) in astrocytes, IL-1-induced YKL-40 expression is p65/p50 independent, and instead regulated by the RelB/p50 complexes. Interestingly, OSM promotes formation of p50/RelB complexes. In conclusion, we found that YKL-40 is up regulated by major pro-inflammatory cytokines during inflammation associated with various disease models. Thus, we uncovered the regulatory mechanism that governs the expression of YKL-40 during sterile inflammation. Next, I studied the role of YKL-40 in GBM in vivo. The major obstacle for treatment of GBM is the unique ability of GBM cells to diffusely infiltrate healthy brain tissue. GBM tumors express high levels of YKL-40, and we showed that administration of YKL-40 increases migration and invasion of U1242 cells in vitro. We used the U1242 cell line for our studies, due to its unique ability to form infiltrative tumors, which reflects the real brain pathology in GBM. Additionally, knock-down of YKL-40 suppresses the migration and invasion of GBM cells in vitro. To study this phenomenon in vivo, we utilized an inducible lentiviral vector containing shRNA to YKL-40.…
Advisors/Committee Members: Tomasz Kordula.
Subjects/Keywords: Biochemistry, Biophysics, and Structural Biology; Life Sciences
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❌
APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
Bhardwaj, R. (2014). REGULATION OF YKL-40 IN STERILE INFLAMMATION AND ITS ROLE IN GLIOBLASTOMA IN VIVO. (Doctoral Dissertation). Virginia Commonwealth University. Retrieved from https://doi.org/10.25772/4JHN-WG10 ; https://scholarscompass.vcu.edu/etd/629
Chicago Manual of Style (16th Edition):
Bhardwaj, Reetika. “REGULATION OF YKL-40 IN STERILE INFLAMMATION AND ITS ROLE IN GLIOBLASTOMA IN VIVO.” 2014. Doctoral Dissertation, Virginia Commonwealth University. Accessed March 04, 2021.
https://doi.org/10.25772/4JHN-WG10 ; https://scholarscompass.vcu.edu/etd/629.
MLA Handbook (7th Edition):
Bhardwaj, Reetika. “REGULATION OF YKL-40 IN STERILE INFLAMMATION AND ITS ROLE IN GLIOBLASTOMA IN VIVO.” 2014. Web. 04 Mar 2021.
Vancouver:
Bhardwaj R. REGULATION OF YKL-40 IN STERILE INFLAMMATION AND ITS ROLE IN GLIOBLASTOMA IN VIVO. [Internet] [Doctoral dissertation]. Virginia Commonwealth University; 2014. [cited 2021 Mar 04].
Available from: https://doi.org/10.25772/4JHN-WG10 ; https://scholarscompass.vcu.edu/etd/629.
Council of Science Editors:
Bhardwaj R. REGULATION OF YKL-40 IN STERILE INFLAMMATION AND ITS ROLE IN GLIOBLASTOMA IN VIVO. [Doctoral Dissertation]. Virginia Commonwealth University; 2014. Available from: https://doi.org/10.25772/4JHN-WG10 ; https://scholarscompass.vcu.edu/etd/629

Virginia Commonwealth University
24.
Yuan, Fang.
DIFFERENTIAL EFFECTS OF GROWTH FACTORS ON GLYCOLYSIS IN OVARIAN CANCER CELLS.
Degree: MS, Biochemistry, 2013, Virginia Commonwealth University
URL: https://doi.org/10.25772/F3JD-BD39
;
https://scholarscompass.vcu.edu/etd/466
► Lysophosphatidic acid (LPA), a naturally-occurring, simple phospholipid, is present at elevated levels in the blood and ascites of ovarian cancer patients. LPA is a ligand…
(more)
▼ Lysophosphatidic acid (LPA), a naturally-occurring, simple phospholipid, is present at elevated levels in the blood and ascites of ovarian cancer patients. LPA is a ligand of seven cell surface G protein-coupled receptors. It has been known as an oncogenic growth factor in ovarian cancer and other types of human malignancies. However, the precise biological functions of LPA in ovarian oncogenesis remain to be fully elucidated. Our laboratory is interested in studying the potential role of LPA, as a tumor microenvironment factor, in regulation of cancer cell metabolism. A fundamental change associated with most cancer is the switch of glucose metabolism from mitochondrial oxidative phosphorylation to aerobic glycolysis, a phenomenon described by Otto Warburg nearly a century ago. This seems to be necessary to meet bioenergetic and biosynthetic demands of rapidly dividing tumor cells. However, the mechanism underlying the switch from aerobic respiration to aerobic glycolysis in cancer cells remains poorly understood.
In this thesis project, my goal was to explore the effect of LPA on glycolysis and to compare LPA with other important growth factors in their capability to promote the glycolytic pathway in ovarian cancer cells. We demonstrated that LPA stimulated aerobic glycolysis as well as cell proliferation in ovarian cancer cell lines. The two parallel responses were LPA dose dependent. To determine whether LPA is unique in driving glycolysis, we compared the effect of LPA with other growth factors, including EGF, insulin and IGF-1 which are all involved in pathogenesis of ovarian cancer. While doses of these growth factors could be adjusted to achieve similar levels of cell proliferation, LPA and EGF were much more potent than insulin and IGF-1 in stimulation of glycolytic flux and lactate production. Therefore, we identified LPA and EGF as highly glycolytic factors relevant to the development and maintenance of the glycolytic phenotype of ovarian cancer cells.
The next part of my study was focused on the molecular mechanism for the differential effects of LPA, EGF, insulin and IGF-1 on glycolytic metabolism. We used the glucose metabolism RT-PCR array to profile expression of glycolytic genes. The most remarkable change induced by LPA and EGF was the robust induction of hexokinase 2 (HK2) that stimulates irreversible entry of glucose to the glycolytic pathway. However, insulin and IGF-1 only weakly induced HK2 expression. Further experimental evidence using HK2 inhibitors indicated that HK2 up-regulation was the critical mediator of LPA-induced glycolysis. Further, the cells grown in LPA and EGF-stimulated conditions appear to show larger volume compared to insulin and IGF-1-treated cells, consistent with the hypothesis that active glycolysis contributes to biosynthetic processes to maintain cell sizes.
Taken together, these findings of the current study revealed high glycolytic effects of LPA and EGF in ovarian cancer and the underlying HK2-mediated mechanism that distinguishes LPA and EGF from other growth…
Advisors/Committee Members: Xianjun Fang.
Subjects/Keywords: Biochemistry, Biophysics, and Structural Biology; Life Sciences
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APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
Yuan, F. (2013). DIFFERENTIAL EFFECTS OF GROWTH FACTORS ON GLYCOLYSIS IN OVARIAN CANCER CELLS. (Thesis). Virginia Commonwealth University. Retrieved from https://doi.org/10.25772/F3JD-BD39 ; https://scholarscompass.vcu.edu/etd/466
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):
Yuan, Fang. “DIFFERENTIAL EFFECTS OF GROWTH FACTORS ON GLYCOLYSIS IN OVARIAN CANCER CELLS.” 2013. Thesis, Virginia Commonwealth University. Accessed March 04, 2021.
https://doi.org/10.25772/F3JD-BD39 ; https://scholarscompass.vcu.edu/etd/466.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Yuan, Fang. “DIFFERENTIAL EFFECTS OF GROWTH FACTORS ON GLYCOLYSIS IN OVARIAN CANCER CELLS.” 2013. Web. 04 Mar 2021.
Vancouver:
Yuan F. DIFFERENTIAL EFFECTS OF GROWTH FACTORS ON GLYCOLYSIS IN OVARIAN CANCER CELLS. [Internet] [Thesis]. Virginia Commonwealth University; 2013. [cited 2021 Mar 04].
Available from: https://doi.org/10.25772/F3JD-BD39 ; https://scholarscompass.vcu.edu/etd/466.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Yuan F. DIFFERENTIAL EFFECTS OF GROWTH FACTORS ON GLYCOLYSIS IN OVARIAN CANCER CELLS. [Thesis]. Virginia Commonwealth University; 2013. Available from: https://doi.org/10.25772/F3JD-BD39 ; https://scholarscompass.vcu.edu/etd/466
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Virginia Commonwealth University
25.
Marion, James D., Jr.
THE ACTIVATION, RECEPTOR COMPLEXING AND ENDOGENOUS REGULATION OF
THE TYPE-I INTERFERON RESPONSE AS IT PERTAINS TO INNATE IMMUNITY.
Degree: PhD, Biochemistry, 2013, Virginia Commonwealth University
URL: https://doi.org/10.25772/C0T2-F683
;
https://scholarscompass.vcu.edu/etd/487
► To defend against pathogen challenge, multi-cellular organisms mount an immune response that recognizes, sequesters and eradicates invading infectious agents. Critical to this safeguard is the…
(more)
▼ To defend against pathogen challenge, multi-cellular organisms mount an immune response that recognizes, sequesters and eradicates invading infectious agents. Critical to this safeguard is the receptor-mediated detection of pathogens. Pathogen recognition then initiates a variety of signaling cascades that lead to the modulation of genes orchestrating an immune response. Toll-like receptor 3 (TLR3), a transmembrane receptor found in endosomes, is vital to the innate immune response against viruses. Double-stranded RNA (dsRNA) stimulation of TLR3 initiates a signaling cascade that leads to the production of type-I interferons and proinflammatory cytokines necessary to trigger the protective defenses of the immune system. Critical to this pathway is the activation of a kinase, TANK binding kinase 1 (TBK1), which phosphorylates the downstream transcription factors, IRF3 and IRF7, and leads to the production of IFN-beta. Interestingly, TBK1 function has been implicated in a number of other signaling cascades ranging from the insulin response and vesicle transport to xenophagy and anti-viral immunity. Increasingly, however, TBK1 dysregulation has been linked to autoimmune disorders and cancers, heightening the need to understand regulatory controls of TBK1. As a result, this dissertation investigates three components of the TLR3 signaling cascade in an attempt to further advance our understanding of the innate immune response. First, investigations into the adjuvant potential of dsRNA reveal that a 139bp dsRNA molecule is a viable candidate for vaccine adjuvant studies. Next,
structural and functional studies of TLR3 with neutralizing antibodies provide evidence for a new TLR-signaling model in which dsRNA:TLR3 signaling units laterally cluster to achieve efficient signaling. Finally, cell-based assays, biophysical experiments and kinetic investigations into the mechanism by which an endogenous regulator of the TLR3 response, SIKE, functions, reveal that SIKE not only inhibits TBK1-mediated IRF3 phosphorylation, but is also a high affinity substrate. Findings from this study further suggest that SIKE regulates a critical catalytic hub not by direct repression of activity, but by redirection of catalysis through substrate affinity. Taken together, the results presented in this dissertation establish a foundation for building long-term studies on the function, regulation and viral subversion of the innate immune response.
Advisors/Committee Members: Jessica K. Bell.
Subjects/Keywords: Biochemistry, Biophysics, and Structural Biology; Life Sciences
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APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
Marion, James D., J. (2013). THE ACTIVATION, RECEPTOR COMPLEXING AND ENDOGENOUS REGULATION OF
THE TYPE-I INTERFERON RESPONSE AS IT PERTAINS TO INNATE IMMUNITY. (Doctoral Dissertation). Virginia Commonwealth University. Retrieved from https://doi.org/10.25772/C0T2-F683 ; https://scholarscompass.vcu.edu/etd/487
Chicago Manual of Style (16th Edition):
Marion, James D., Jr. “THE ACTIVATION, RECEPTOR COMPLEXING AND ENDOGENOUS REGULATION OF
THE TYPE-I INTERFERON RESPONSE AS IT PERTAINS TO INNATE IMMUNITY.” 2013. Doctoral Dissertation, Virginia Commonwealth University. Accessed March 04, 2021.
https://doi.org/10.25772/C0T2-F683 ; https://scholarscompass.vcu.edu/etd/487.
MLA Handbook (7th Edition):
Marion, James D., Jr. “THE ACTIVATION, RECEPTOR COMPLEXING AND ENDOGENOUS REGULATION OF
THE TYPE-I INTERFERON RESPONSE AS IT PERTAINS TO INNATE IMMUNITY.” 2013. Web. 04 Mar 2021.
Vancouver:
Marion, James D. J. THE ACTIVATION, RECEPTOR COMPLEXING AND ENDOGENOUS REGULATION OF
THE TYPE-I INTERFERON RESPONSE AS IT PERTAINS TO INNATE IMMUNITY. [Internet] [Doctoral dissertation]. Virginia Commonwealth University; 2013. [cited 2021 Mar 04].
Available from: https://doi.org/10.25772/C0T2-F683 ; https://scholarscompass.vcu.edu/etd/487.
Council of Science Editors:
Marion, James D. J. THE ACTIVATION, RECEPTOR COMPLEXING AND ENDOGENOUS REGULATION OF
THE TYPE-I INTERFERON RESPONSE AS IT PERTAINS TO INNATE IMMUNITY. [Doctoral Dissertation]. Virginia Commonwealth University; 2013. Available from: https://doi.org/10.25772/C0T2-F683 ; https://scholarscompass.vcu.edu/etd/487

Western Michigan University
26.
Adiyodi Veetil, Sandhya N.
Glucosamine-Induced Insulin Resistance in Primary Rat Hepatocytes and the Role of Selenium as an Insulin Mimetic.
Degree: PhD, Chemistry, 2011, Western Michigan University
URL: https://scholarworks.wmich.edu/dissertations/475
► Type 2 diabetes is mediated by insulin resistance, the inability of insulin to elicit a normal biological response in insulin responsive tissues. Several cellular…
(more)
▼ Type 2 diabetes is mediated by insulin resistance, the inability of insulin to elicit a normal biological response in insulin responsive tissues. Several cellular models have been utilized to determine the mechanism of induction of insulin resistance but questions remain unanswered. One model, implicates the products of the Hexosamine Biosynthetic Pathway (HBP) in the induction of insulin resistance under hyperglycemia. The major end product of HBP, UDP-GlcNAc, is the substrate for O-GlcNAc transferase, an enzyme that catalyzes the O-GlcNAcylation of numerous proteins. This modification may play a role in induction of insulin resistance and thus needs to be evaluated in different cell types. Therefore, we set out to test whether or not insulin resistance could be established in primary rat hepatocytes. To accomplish this we used a HBP precursor, glucosamine and first assessed whether or not insulin resistance was established by evaluating insulin's effect on a key signaling protein, Akt.
Results indicate that the insulin induced phosphorylation of Akt was decreased in the presence of glucosamine when compared to the control, suggesting an insulin resistant state. Signal protein activation is very important for the insulin regulation of gene expression. If the activation of signal proteins is altered, then the effect on gene expression should also be altered. Results show that under glucosamine treatment, insulin was no longer able to control the gene expression of a number of key enzymes in major metabolic pathways. Additionally an increase in O-GlcNAc modified proteins under glucosamine compared to the control was observed and a number of these proteins were identified through LC-MS.
Lastly, the effect of selenium, an insulin mimetic agent, was examined in this model system. The effects of selenium on the phosphorylation of the insulin signaling protein, Akt and on the expression of the key enzymes of the major metabolic pathways both under normal and insulin resistant conditions were examined. Results show that selenium is a potent insulin mimetic not only under the normal/control condition, but also under the insulin resistant condition as well.
Advisors/Committee Members: Dr. Susan R. Stapleton, Dr. David S. Reinhold, Dr. Pamela Hoppe.
Subjects/Keywords: Biochemistry, Biophysics, and Structural Biology; Chemistry
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❌
APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
Adiyodi Veetil, S. N. (2011). Glucosamine-Induced Insulin Resistance in Primary Rat Hepatocytes and the Role of Selenium as an Insulin Mimetic. (Doctoral Dissertation). Western Michigan University. Retrieved from https://scholarworks.wmich.edu/dissertations/475
Chicago Manual of Style (16th Edition):
Adiyodi Veetil, Sandhya N. “Glucosamine-Induced Insulin Resistance in Primary Rat Hepatocytes and the Role of Selenium as an Insulin Mimetic.” 2011. Doctoral Dissertation, Western Michigan University. Accessed March 04, 2021.
https://scholarworks.wmich.edu/dissertations/475.
MLA Handbook (7th Edition):
Adiyodi Veetil, Sandhya N. “Glucosamine-Induced Insulin Resistance in Primary Rat Hepatocytes and the Role of Selenium as an Insulin Mimetic.” 2011. Web. 04 Mar 2021.
Vancouver:
Adiyodi Veetil SN. Glucosamine-Induced Insulin Resistance in Primary Rat Hepatocytes and the Role of Selenium as an Insulin Mimetic. [Internet] [Doctoral dissertation]. Western Michigan University; 2011. [cited 2021 Mar 04].
Available from: https://scholarworks.wmich.edu/dissertations/475.
Council of Science Editors:
Adiyodi Veetil SN. Glucosamine-Induced Insulin Resistance in Primary Rat Hepatocytes and the Role of Selenium as an Insulin Mimetic. [Doctoral Dissertation]. Western Michigan University; 2011. Available from: https://scholarworks.wmich.edu/dissertations/475

University of Iowa
27.
Zhu, Yueming.
Polychlorinated biphenyl (PCB)-induced oxidative stress mediates cytotoxicity in human breast and prostate epithelial cells.
Degree: PhD, Free Radical and Radiation Biology, 2011, University of Iowa
URL: https://ir.uiowa.edu/etd/3026
► This thesis describes studies that are designed to investigates the hypothesis that <em>mitochondrial production of reactive oxygen species (O2*- and H2O2) cause oxidative stress…
(more)
▼ This thesis describes studies that are designed to investigates the hypothesis that
<em>mitochondrial production of reactive oxygen species
(O
2*- and
H
2O
2) cause oxidative stress during PCB
exposure and this increased production of ROS contributes to the biological effects of
PCBs on cell proliferation in human breast and prostate epithelial cells.</em> Exponentially growing non-malignant human breast epithelial cells (MCF-10A) and
non-malignant human prostate epithelial cells (RWPE-1) were treated with selected PCBs
and their metabolites (PCB3, 77, 153, Aroclor and 4ClBQ). Results showed that PCBs and
their metabolites could significantly inhibit MCF-10A and RWPE-1 cell growth as well as
inducing clonogenic cell killing. These PCBs were also found to increase steady-state
levels of mitochondrial O
2*- and
H
2O
2. Furthermore, the same PCBs were also
found to induce alterations in SOD activities in MCF-10A and RWPE-1 cells. Finally,
treatment with either N-acetyl-cysteine (NAC), or the combination of polyethylene glycol
(PEG) conjugated CuZnSOD and PEG-catalase added 1 hour after PCBs, significantly
protected MCF-10A and RWPE-1 cells from PCB-induced toxicity even when added following
PCB exposure. Similar experiments were also accomplished using airborne PCBs treated RWPE-1 cells.
4-OH-PCB11, a metabolite of airborne PCB 11 is shown to lead to steady-state increases
in superoxide and hydroperoxides in exponentially growing RWPE-1 human nonmalignant
prostate epithelial cells. This increased level of ROS was accompanied by the inhibition
of cell growth and clonogenic cell killing. Furthermore treatment of cells with
antioxidants one hour following exposure to 4-OH-PCB11 was able to significantly
diminish the toxicity in human prostate epithelial cells. These results strongly
supported the hypothesis that exposure to PCBs or their metabolites can induce the
cytotoxicity and alterations in cellular proliferation as well as causing oxidative
stress in exponentially growing human breast and prostate epithelial cells. More
importantly, the data also provide clear evidence that antioxidant manipulations after
PCB exposure are capable of protecting human cells against PCB-induced cytotoxicity.
Based on these observations, the long term goal of this work is to develop a mechanism
based biochemical rationale for the development of pharmaceutical manipulations to
protect humans from PCB intoxication.
Advisors/Committee Members: Spitz, Douglas Robert (supervisor).
Subjects/Keywords: Other Biochemistry; Biophysics; and Structural Biology
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❌
APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
Zhu, Y. (2011). Polychlorinated biphenyl (PCB)-induced oxidative stress mediates cytotoxicity in human breast and prostate epithelial cells. (Doctoral Dissertation). University of Iowa. Retrieved from https://ir.uiowa.edu/etd/3026
Chicago Manual of Style (16th Edition):
Zhu, Yueming. “Polychlorinated biphenyl (PCB)-induced oxidative stress mediates cytotoxicity in human breast and prostate epithelial cells.” 2011. Doctoral Dissertation, University of Iowa. Accessed March 04, 2021.
https://ir.uiowa.edu/etd/3026.
MLA Handbook (7th Edition):
Zhu, Yueming. “Polychlorinated biphenyl (PCB)-induced oxidative stress mediates cytotoxicity in human breast and prostate epithelial cells.” 2011. Web. 04 Mar 2021.
Vancouver:
Zhu Y. Polychlorinated biphenyl (PCB)-induced oxidative stress mediates cytotoxicity in human breast and prostate epithelial cells. [Internet] [Doctoral dissertation]. University of Iowa; 2011. [cited 2021 Mar 04].
Available from: https://ir.uiowa.edu/etd/3026.
Council of Science Editors:
Zhu Y. Polychlorinated biphenyl (PCB)-induced oxidative stress mediates cytotoxicity in human breast and prostate epithelial cells. [Doctoral Dissertation]. University of Iowa; 2011. Available from: https://ir.uiowa.edu/etd/3026

University of Pennsylvania
28.
Zhang, Yao.
Exploring the World of Helix Association: Disease Mechanism, Basic Folding and Novel Design.
Degree: 2011, University of Pennsylvania
URL: https://repository.upenn.edu/edissertations/973
► Helix association provides an efficient model for studying the fundamental principles behind protein folding. It also serves as a suitable template for the design of…
(more)
▼ Helix association provides an efficient model for studying the fundamental principles behind protein folding. It also serves as a suitable template for the design of proteins with novel functions. This thesis begins by investigating the role of transmembrane helix association in protein folding, where a novel “protein-folding-centric” viral fusion model has been proposed here to explain the membrane-fusion process of paramyxovirus. Furthermore, the forces driving membrane helix association, which determine both affinity and orientation, have been quantitatively studied using a model membrane peptide MS1. Finally, two examples are discussed that illustrate the application of helix association in novel protein design. A pH-switchable drug delivery system for the endosomal escape of biomacromolecular therapeutics has been designed using the helix-association model. The sequence is designed to form a stable water-soluble helix bundle at pH 7.4 and to insert in membrane at lower pH to promote endosomal escape. The most successful sequence shows selective release for biomacromolecule (ATP and miRNA) at lower pH (pH 5.4). The assembly of the designed peptide has been studied in aqueous buffer, detergent micelle and model lipid bilayer using the most successful sequence. Also, the paradigm of helix association has been applied to the design of a membrane metalloprotein, which can serve as a template for further design of membrane metalloenzymes. In summary, the work in this thesis has established an efficient model for helix association that can be used to solve problems in both basic and applied research.
Subjects/Keywords: helix assocaiton; Biochemistry, Biophysics, and Structural Biology
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❌
APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
Zhang, Y. (2011). Exploring the World of Helix Association: Disease Mechanism, Basic Folding and Novel Design. (Thesis). University of Pennsylvania. Retrieved from https://repository.upenn.edu/edissertations/973
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):
Zhang, Yao. “Exploring the World of Helix Association: Disease Mechanism, Basic Folding and Novel Design.” 2011. Thesis, University of Pennsylvania. Accessed March 04, 2021.
https://repository.upenn.edu/edissertations/973.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
MLA Handbook (7th Edition):
Zhang, Yao. “Exploring the World of Helix Association: Disease Mechanism, Basic Folding and Novel Design.” 2011. Web. 04 Mar 2021.
Vancouver:
Zhang Y. Exploring the World of Helix Association: Disease Mechanism, Basic Folding and Novel Design. [Internet] [Thesis]. University of Pennsylvania; 2011. [cited 2021 Mar 04].
Available from: https://repository.upenn.edu/edissertations/973.
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation
Council of Science Editors:
Zhang Y. Exploring the World of Helix Association: Disease Mechanism, Basic Folding and Novel Design. [Thesis]. University of Pennsylvania; 2011. Available from: https://repository.upenn.edu/edissertations/973
Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

University of Washington
29.
Vittal, Vinayak.
Structural and Functional Characterization of Non-Canonical Ubiquitin Conjugating Enzymes.
Degree: PhD, 2015, University of Washington
URL: http://hdl.handle.net/1773/33103
► Thirty-five years ago, during its initial discovery, the field of ubiquitination could not have been more aptly named. In fact, as the study of ubiquitin…
(more)
▼ Thirty-five years ago, during its initial discovery, the field of ubiquitination could not have been more aptly named. In fact, as the study of ubiquitin (Ub) expands, it is becoming clear that this small post-translational modification is far more ubiquitous than the founders of this field could have originally imagined. Initially identified as a protein degradation signal, Ub is now known to regulate vastly disparate cellular functions, including the DNA-damage response and immune signaling. This small (8.5kDa) modification achieves its diverse signaling capabilities partly through the enzymes that catalyze its attachment to cellular substrates. These enzymes (E1, E2, and E3) work in succession to identify and modify substrates with varying Ub signals. The manner in which Ub is attached (monoubiquition or 8 types of polyubiquitin chains) and the site of attachment dictate a substrate's cellular fate. Furthermore, much of the diversity in Ub signaling can be attributed to the
structural characteristics of the enzymes in the ubiquitin cascade. This thesis examines the
structural and biochemical features of two E2 ubiquitin-conjugating enzymes, Ube2w and Ube2h. First, we show that Ube2w specifically attaches monoubiquitin to the N-terimni of disordered protein substrates. To attach this unique Ub modification, Ube2w has evolved a non-canonical domain architecture, solved by our group using nuclear magnetic resonance (NMR) techniques. We also show, that another E2, Ube2h, contains a disordered C-terminus and that it can ubiquitinate a substrate, histone H2A/H2B, in vitro, in the absence of an E3. Studies presented here build the foundations to understand the cellular impact of Ube2w and Ube2h.
Advisors/Committee Members: Klevit, Rachel E (advisor).
Subjects/Keywords: biochemistry; biology; biophysics; structural; ubiquitin; Biochemistry; Biophysics; biological chemistry
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APA ·
Chicago ·
MLA ·
Vancouver ·
CSE |
Export
to Zotero / EndNote / Reference
Manager
APA (6th Edition):
Vittal, V. (2015). Structural and Functional Characterization of Non-Canonical Ubiquitin Conjugating Enzymes. (Doctoral Dissertation). University of Washington. Retrieved from http://hdl.handle.net/1773/33103
Chicago Manual of Style (16th Edition):
Vittal, Vinayak. “Structural and Functional Characterization of Non-Canonical Ubiquitin Conjugating Enzymes.” 2015. Doctoral Dissertation, University of Washington. Accessed March 04, 2021.
http://hdl.handle.net/1773/33103.
MLA Handbook (7th Edition):
Vittal, Vinayak. “Structural and Functional Characterization of Non-Canonical Ubiquitin Conjugating Enzymes.” 2015. Web. 04 Mar 2021.
Vancouver:
Vittal V. Structural and Functional Characterization of Non-Canonical Ubiquitin Conjugating Enzymes. [Internet] [Doctoral dissertation]. University of Washington; 2015. [cited 2021 Mar 04].
Available from: http://hdl.handle.net/1773/33103.
Council of Science Editors:
Vittal V. Structural and Functional Characterization of Non-Canonical Ubiquitin Conjugating Enzymes. [Doctoral Dissertation]. University of Washington; 2015. Available from: http://hdl.handle.net/1773/33103

McMaster University
30.
Stalker, Leanne.
CHARACTERIZING THE FUNCTION AND REGULATORY MECHANISMS OF THE HISTONE DEMETHYLASE KDM5B: INSIGHTS INTO THE COMPLEXITY OF EPIGENETIC REGULATION.
Degree: PhD, 2013, McMaster University
URL: http://hdl.handle.net/11375/12920
► KDM5b acts as a transcriptional repressor through its ability to demethylate tri-methylated lysine (K) 4 on histone H3 (H3K4me3). Demethylation of this histone modification…
(more)
▼ KDM5b acts as a transcriptional repressor through its ability to demethylate tri-methylated lysine (K) 4 on histone H3 (H3K4me3). Demethylation of this histone modification leads to transcriptional repression and downstream biological effects on gene expression. KDM5b is involved in the regulation of differentiation and can exert an oncogenic and a tumour suppressive role depending on cellular context, making it an attractive future target for pharmaceutical intervention. Work from our group has shown that KDM5b expression is linked to differentiation, and that recruitment of the enzyme does not always result in an alteration of H3K4me3. Additionally, work from our group, as well as others, has failed to observe H3K4me3 demethylation by KDM5b in nucleosomal preparations. We therefore hypothesized that KDM5b may exert its demethylase potential on alternative histone targets and that KDM5b requires enzymatic co-factors to demethylate nucleosomes, similar to what is observed for other histone-modifying proteins. In this thesis, we describe KDM5b as having an alternate histone target, di-methylated histone H2B lysine 43 (H2BK43me2). We show that this methyl mark is the primary target for KDM5b, and that the expression level of H2BK43me2 is directly related to the process of differentiation. We additionally present a novel co-factor for KDM5b, the co-repressor TLE4 of the Groucho/TLE family. The presence of TLE4 is required and sufficient to confer nucleosomal demethylase activity to KDM5b, a novel discovery for any of the KDM5 family members. Overall, this work has described both an additional KDM5b target, and detailed requirements for KDM5b nucleosomal demethylation, advancing our understanding of how this enzyme is regulated in vivo. The novel aspects of KDM5b regulation presented within this thesis provide a framework from which future studies can be designed. This work contributes to our overall understanding of epigenetic regulation and will potentially aid in the development of novel anti-cancer therapeutic strategies.
Doctor of Philosophy (PhD)
Advisors/Committee Members: Truant, Ray, Biochemistry.
Subjects/Keywords: demethylase; histone; KDM5; epigenetics; Biochemistry, Biophysics, and Structural Biology; Biochemistry, Biophysics, and Structural Biology
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CSE |
Export
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Manager
APA (6th Edition):
Stalker, L. (2013). CHARACTERIZING THE FUNCTION AND REGULATORY MECHANISMS OF THE HISTONE DEMETHYLASE KDM5B: INSIGHTS INTO THE COMPLEXITY OF EPIGENETIC REGULATION. (Doctoral Dissertation). McMaster University. Retrieved from http://hdl.handle.net/11375/12920
Chicago Manual of Style (16th Edition):
Stalker, Leanne. “CHARACTERIZING THE FUNCTION AND REGULATORY MECHANISMS OF THE HISTONE DEMETHYLASE KDM5B: INSIGHTS INTO THE COMPLEXITY OF EPIGENETIC REGULATION.” 2013. Doctoral Dissertation, McMaster University. Accessed March 04, 2021.
http://hdl.handle.net/11375/12920.
MLA Handbook (7th Edition):
Stalker, Leanne. “CHARACTERIZING THE FUNCTION AND REGULATORY MECHANISMS OF THE HISTONE DEMETHYLASE KDM5B: INSIGHTS INTO THE COMPLEXITY OF EPIGENETIC REGULATION.” 2013. Web. 04 Mar 2021.
Vancouver:
Stalker L. CHARACTERIZING THE FUNCTION AND REGULATORY MECHANISMS OF THE HISTONE DEMETHYLASE KDM5B: INSIGHTS INTO THE COMPLEXITY OF EPIGENETIC REGULATION. [Internet] [Doctoral dissertation]. McMaster University; 2013. [cited 2021 Mar 04].
Available from: http://hdl.handle.net/11375/12920.
Council of Science Editors:
Stalker L. CHARACTERIZING THE FUNCTION AND REGULATORY MECHANISMS OF THE HISTONE DEMETHYLASE KDM5B: INSIGHTS INTO THE COMPLEXITY OF EPIGENETIC REGULATION. [Doctoral Dissertation]. McMaster University; 2013. Available from: http://hdl.handle.net/11375/12920
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