Biochemical investigations of protein arginine N-methyltransferase 2
department or school: Pharmaceutical Sciences, Faculty of
data provider: DSpace
degree campus: UBCV
creator: Rowley, Michael James Anthony
issued: 2024-05-09T17:53:19Z
issued: 2024
related degree: Doctor of Philosophy - PhD
degree grantor: University of British Columbia
description: Arginine methylation is a prevalent post-translational modification in biology present on 0.5% of total arginine proteins in mammalian cells. Protein arginine methylation is a key epigenetic modification on histone proteins, that can promote or inhibit gene expression depending on the type and site of arginine methylation. Protein arginine N-methyltransferases (PRMTs) are the epigenetic writers responsible for arginine methylation and have emerged as therapeutic targets in several health disorders including neurodegenerative and cardiovascular disease, and cancer. PRMT2 was recently added to the list of epigenetic writers associated with oncogenic transcriptional programming, specifically in glioblastoma (GBM), hepatocellular carcinoma (HCC), and renal cell carcinoma (RCC). Unlike the other PRMT enzymes, the function of PRMT2 in cells remains enigmatic due to its lackluster in vitro methylation kinetics, making it difficult to characterize. The purpose of this research is to develop techniques for measuring PRMT2 interactions and to use these techniques to characterize how PRMT2 interacts with other proteins and histones. The development of the filter binding and phosphor screening (FBAPS) assay improved the throughput of the traditional filter binding assay used to characterize kinetic parameters of PRMT enzymes. Characterization of PRMT2 through filter binding assay revealed that exposing the radiolabeled S-adenosyl-L-methionine (SAM) cofactor to elevated temperatures for long periods of time create degradation products that confound analysis. We used differential scanning fluorimetry (DSF) to characterize PRMT2 thermal stability to identify conditions that may improve PRMT2 catalytic activity and to measure PRMT2 binding interactions. We identified several excipients that could thermally stabilize PRMT2 but did not improve catalytic activity. Additionally, we found that PRMT2 can interact with SAM and sinefungin (a SAM analogue) in a thermally stabilizing manner.…
aggregated CHO: https://circle.library.ubc.ca/rest/handle/2429/88263
note: BIOCHEMICAL INVESTIGATIONS OF PROTEIN ARGININE N-METHYLTRANSFERASE 2 by Michael James Anthony Rowley B.Sc. (Hons), The University of Calgary, 2019 A THESIS SUBMITTED IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF DOCTOR OF PHILOSOPHY in THE FACULTY OF GRADUATE AND POSTDOCTORAL STUDIES (Pharmaceutical Sciences) THE UNIVERSITY OF BRITISH COLUMBIA (Vancouver) May 2024 © Michael James Anthony Rowley, 2024 ii The following individuals certify that they have read, and recommend to the Faculty of Graduate and Postdoctoral Studies for acceptance, the dissertation entitled: BIOCHEMICAL INVESTIGATIONS OF PROTEIN ARGININE N-METHYLTRANSFERASE 2 submitted by Michael James Anthony Rowley in partial fulfilment of the requirements for the degree of Doctor of Philosophy in Pharmaceutical Sciences Examining Committee: Dr. Adam Frankel, Associate Professor, Pharmaceutical Sciences, UBC Supervisor Dr. Filip van Petegem, Professor, Biochemistry and Molecular Biology, UBC Supervisory Committee Member Dr. Martin Tanner, Professor, Chemistry, UBC University Examiner Dr. Judy Wong, Professor, Pharmaceutical Sciences, UBC University Examiner Dr. Muriel le Romancer, Professor, Centre de Recherche en Cancérologie de Lyon, Université de Lyon External Examiner Additional Supervisory Committee Members: Dr. Brent Page, Assistant Professor, Pharmaceutical Sciences, UBC Supervisory Committee Member Dr. Karla Williams, Associate Professor, Pharmaceutical Sciences, UBC Supervisory Committee Member iii Abstract Arginine methylation is a prevalent post-translational modification in biology present on 0.5% of total arginine proteins in mammalian cells. Protein arginine methylation is a key epigenetic modification on histone proteins, that can promote or inhibit gene expression depending on the type and site of arginine methylation. Protein arginine N-methyltransferases (PRMTs) are the epigenetic writers responsible for arginine methylation and have emerged as therapeutic targets in several …
has type: Thesis/Dissertation
date issued: 2024-11
is shown at: 10.14288/1.0442425
language: eng
degree discipline: Pharmaceutical Sciences
provider: Vancouver : University of British Columbia Library
publisher: University of British Columbia
rights: Attribution-NonCommercial-NoDerivatives 4.0 International
rights URI: http://creativecommons.org/licenses/by-nc-nd/4.0/
scholar level: Graduate
contributor: Frankel, Adam
type: Text
identifier URI: http://hdl.handle.net/2429/88263