Identifying and quantifying sites of protein methylation by heavy methyl SILAC

Current Protocols in Protein Science
Shao-En Ong, Matthias Mann

Abstract

A new appreciation of protein methylation comes with the recent discovery of demethylases, now placing methylation in the realm of a transient, reversible modification. Classical approaches to study methylation are laborious and involve radioactive, in vitro, enzyme-substrate labeling experiments with purified proteins. Mass spectrometry-based proteomics allows the unbiased analysis of complex protein mixtures and is increasingly applied to the study of post-translational modifications. However, it is particularly challenging to study methylation by proteomics because of the number of residues affected and the degree of methylation that can occur. Heavy methyl SILAC is a metabolic labeling strategy that harnesses the cell's machinery to convert a nonradioactive, stable isotope labeled version of methionine into the 'heavy' biological methyl donor S-adenosylmethionine. Cells incorporate this 'heavy' methyl group throughout their methylated substrates. This technique increases confidence in identifying and quantifying of sites of protein methylation.

References

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Citations

Nov 3, 2009·Journal of Proteome Research·Junmei ZhangYingming Zhao
Nov 16, 2012·Molecular & Cellular Proteomics : MCP·John C FiskLaurie K Read
May 1, 2012·Cell Cycle·Scott M Carlson, Forest M White
May 7, 2013·Epigenetics : Official Journal of the DNA Methylation Society·Xing-Jun CaoBenjamin A Garcia
Nov 2, 2016·Current Protocols in Protein Science·Xing-Jun Cao, Benjamin A Garcia
Nov 3, 2016·Cold Spring Harbor Perspectives in Medicine·Scott M Carlson, Or Gozani
Apr 25, 2020·Frontiers in Cell and Developmental Biology·Ariana KupaiScott B Rothbart
Oct 10, 2008·Analytical and Bioanalytical Chemistry·Anton IliukW Andy Tao
Nov 3, 2015·Current Protocols in Protein Science·Kathrine B Sylvestersen, Michael L Nielsen
Feb 13, 2020·Journal of Proteome Research·Kersti KaruKenji Okuse
Feb 2, 2013·New Biotechnology·Marie Rogne, Kjetil Taskén

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