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Using affinity purification coupled with stable isotope labeling by amino acids in cell culture quantitative mass spectrometry to identify novel interactors/substrates of protein arginine methyltransferases.
- Source :
-
Methods (San Diego, Calif.) [Methods] 2020 Mar 15; Vol. 175, pp. 44-52. Date of Electronic Publication: 2019 Nov 30. - Publication Year :
- 2020
-
Abstract
- The protein arginine methyltransferase family (PRMT) is known as being the catalytic driving force for arginine methylation. This specific type of post translational modification is extensively used in biological processes, and therefore is highly relevant in the pathology of a profusion of diseases. Since altered PRMT expression or deregulation has been shown to contribute to a vast range of those diseases including cancer, their study is of great interest. Although an increasing number of substrates are being discovered for each PRMT, large scale proteomic methods can be used to identify novel interactors/substrates, further elucidating the role that PRMTs perform in physiological or disease states. Here, we describe the use of affinity purification (AP) coupled with stable isotope labeling with amino acids in cell culture (SILAC) quantitative mass spectrometry (MS) to identify protein interactors and substrates of PRMTs. We also explore the possibility of exploiting the fact most PRMTs display lower dissociation rates with their hypomethylated substrates as a strategy to increase the proportion of substrates identified in AP/MS studies.<br />Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.<br /> (Copyright © 2019. Published by Elsevier Inc.)
- Subjects :
- Amino Acids metabolism
Arginine analysis
Arginine chemistry
Arginine metabolism
Gene Expression
Histones chemistry
Histones metabolism
Humans
Isotope Labeling
MCF-7 Cells
Nuclear Proteins genetics
Nuclear Proteins metabolism
Protein Binding drug effects
Protein-Arginine N-Methyltransferases genetics
Protein-Arginine N-Methyltransferases metabolism
Recombinant Proteins
Chromatography, Affinity methods
Enzyme Inhibitors chemistry
Mass Spectrometry methods
Protein-Arginine N-Methyltransferases antagonists & inhibitors
Protein-Arginine N-Methyltransferases chemistry
Proteomics methods
Subjects
Details
- Language :
- English
- ISSN :
- 1095-9130
- Volume :
- 175
- Database :
- MEDLINE
- Journal :
- Methods (San Diego, Calif.)
- Publication Type :
- Academic Journal
- Accession number :
- 31794835
- Full Text :
- https://doi.org/10.1016/j.ymeth.2019.11.015