In vitro methylation by methanol: proteomic screening and prevalence investigation

Analytica Chimica Acta
Guoqiang ChenZhili Li

Abstract

It is assumed that much more functional importance for protein activity than expected may be granted by methylation that occurs at the side-chain of aspartate or glutamate residue. In vitro methylation mainly comes from the use of methanol in sample preparation prior to MS analysis. In this study, we first performed the methylation site-directed proteomic screening of bovine serum albumin, ovalbumin and 20S proteasome for gel staining using a meaningfully indicative MS-pattern of peak tag (termed as 4P tag) and manual inspection for mass spectral data. As a result, there were 17 proteolytic peptides with 20 modified sites confirmed to be in vitro methylated. Subsequently, the prevalence investigation was performed, focusing on the reaction kinetic behavior of in vitro methylation. This study provided a simple and robust approach for confirmation of in vitro methylation by methanol, as well as the precautious guide for the use of methanol in proteomic study.

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Citations

Oct 17, 2012·Proceedings of the National Academy of Sciences of the United States of America·Narkhyun BaeGert Lubec
Nov 26, 2010·Analytical Biochemistry·Laura D R DavisIllimar Altosaar
Oct 22, 2013·Journal of Molecular and Cellular Cardiology·Ziyou CuiAldrin V Gomes
Nov 22, 2012·FEMS Microbiology Reviews·Jerry Eichler, Julie Maupin-Furlow
Oct 5, 2016·Journal of Proteome Research·Katelyn E Caslavka ZempelSteven G Clarke
Jan 17, 2017·Journal of Proteome Research·Melda Z GürayAlan A Doucette
Apr 10, 2017·FEBS Letters·Virginie F RheinJohn E Walker

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