A CHEMOPROTEOMIC TECHNIQUE FOR THE IDENTIFICATION OF PROTEINS REGULATED BY COENZYME A

Nelson, Ornella Daisha

A CHEMOPROTEOMIC TECHNIQUE FOR THE IDENTIFICATION OF PROTEINS REGULATED BY COENZYME A

dc.contributor.author	Nelson, Ornella Daisha
dc.contributor.chair	Lin, Hening
dc.contributor.committeeMember	Crane, Brian
dc.contributor.committeeMember	Aye, Yimon
dc.date.accessioned	2019-04-02T14:00:35Z
dc.date.available	2021-01-02T07:01:00Z
dc.date.issued	2018-12-30
dc.description	Supplemental file(s) description: APPENDIX B - Dataset B2.1, APPENDIX B - Dataset B2.2
dc.description.abstract	Coenzyme A (CoA) is found in all living organisms and is involved in numerous cellular biochemical reactions. CoA and its derivatives function as cofactors in many cellular pathways and are involved in protein regulation through post-translational modifications as well as competitive and allosteric interactions. Deregulation of CoA has implications in various human diseases including cancer, diabetes and neurodegeneration. Given the importance of CoA, I synthesized a biotin-tagged CoA probe to identify CoA-binding proteins. Interestingly, my chemical proteomic approach identified several metabolic enzymes that are not known to utilize CoA. I biochemically validated the top hit, mitochondrial malic enzyme 2 (ME2), as a CoA-binding protein. ME2 is a NAD(P)-dependent enzyme that converts malate to pyruvate and at the same time generates NAD(P)H. ME2 plays an important role in numerous cellular processes including: proliferation, insulin secretion, senescence, glutamine metabolism and reactive oxygen species (ROS) homeostasis. I found that ME2 binds CoA tightly and, through mutation studies, identified the CoA-binding site of ME2. By screening various CoA species, I discovered a specific CoA molecule can dramatically increase ME2 enzymatic activity. Furthermore, I determined that the identified CoA molecule promotes ME2 tetramer formation and may have a role in relieving oxidative stress. This work demonstrates that cofactor-based chemical proteomic strategies work well to identify novel protein interactions. Expansion of this technique may prove useful in better understanding cellular regulation.
dc.identifier.doi	https://doi.org/10.7298/vwww-cv58
dc.identifier.other	Nelson_cornellgrad_0058F_11179
dc.identifier.other	http://dissertations.umi.com/cornellgrad:11179
dc.identifier.other	bibid: 10758058
dc.identifier.uri	https://hdl.handle.net/1813/64918
dc.language.iso	en_US
dc.rights	Attribution-NonCommercial-NoDerivatives 4.0 International
dc.rights.uri	https://creativecommons.org/licenses/by-nc-nd/4.0/
dc.subject	Chemoproteomics
dc.subject	Coenzyme A
dc.subject	Desulfotomaculum reducens
dc.subject	Lysine succinylation
dc.subject	Malic enzyme 2
dc.subject	Sirtuin
dc.subject	Molecular biology
dc.subject	Biochemistry
dc.subject	Chemistry
dc.title	A CHEMOPROTEOMIC TECHNIQUE FOR THE IDENTIFICATION OF PROTEINS REGULATED BY COENZYME A
dc.type	dissertation or thesis
dcterms.license	https://hdl.handle.net/1813/59810
thesis.degree.discipline	Chemistry and Chemical Biology
thesis.degree.grantor	Cornell University
thesis.degree.level	Doctor of Philosophy
thesis.degree.name	Ph. D., Chemistry and Chemical Biology