Date Approved
6-5-2024
Embargo Period
6-5-2025
Document Type
Dissertation
Degree Name
Doctor of Philosophy (Ph.D.)
Department
Molecular Biology
College
Rowan-Virtua School of Translational Biomedical Engineering & Sciences
Advisor
Brian Weiser, Ph.D.
Committee Member 1
Randy Strich, Ph.D.
Committee Member 2
Gary S. Goldberg, Ph.D.
Committee Member 3
Mikhail Anikin, Ph.D.
Committee Member 4
Valerie J. Carabetta, Ph.D.
Keywords
deacylase activity; dimerization; drug discovery; SIRT2; small inhibitory molecules
Subject(s)
Clinical Biochemistry; Sirtuins
Disciplines
Biochemistry, Biophysics, and Structural Biology | Life Sciences | Molecular Biology
Abstract
Human sirtuin isoform 2 (SIRT2) is an NAD+-dependent enzyme that removes various acyl modifications from lysine residues. SIRT2 is involved in many biological processes, such as transcription, DNA repair, cell proliferation, and metabolism. Due to its diverse functions, SIRT2 is implicated in various disease states, including cancers and neurodegenerative disorders. This dissertation aimed to understand the regulation of SIRT2’s deacylase activities and to discover small molecule modulators of SIRT2. In this work, we found that SIRT2 dimerized in solution, and the dimerization decreased the deacetylase activities without affecting the defatty-acylase activity of this protein. Dimerized SIRT2 dissociates into monomers upon binding long fatty acylated substrates but not acetylated substrate. In addition, SIRT2 dimerizes in cells, as evidenced by split GFP technology. Furthermore, SIRT2 dimerization can be disrupted genetically by point mutations and pharmacologically by ascorbyl palmitate. Our work also discovered a small molecule (8008-3660) (1) from a novel high-throughput screen that inhibits the enzyme’s deacetylase and defatty-acylase activities with potency comparable to known defatty-acylase inhibitors. Altogether, the genetic and pharmacological modulators discovered in this work have the potential to clarify different SIRT2’s deacylase activities in diseased and healthy cells and provide evidence that SIRT2 can be utilized as a therapeutic target for treating various diseases.
Recommended Citation
Yang, Jie, "DISCOVERY OF OLIGOMERIC STATES AND SMALL MOLECULES THAT AFFECT DEACYLASE ACTIVITIES OF HUMAN SIRTUIN-2 (SIRT2)" (2024). Theses and Dissertations. 3234.
https://rdw.rowan.edu/etd/3234
