Antibody-Free Approach for the Global Analysis of Protein Methylation,Analytical Chemistry

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Antibody-Free Approach for the Global Analysis of Protein Methylation
Analytical Chemistry ( IF 6.7 ) Pub Date : 2016-11-14 00:00:00 , DOI: 10.1021/acs.analchem.6b02872
Keyun Wang _{1,

2} , Mingming Dong _{1,

2} , Jiawei Mao _{1,

2} , Yan Wang _{1,

2} , Yan Jin _{1,

2} , Mingliang Ye _{1,

2} , Hanfa Zou _{1,

2}

Affiliation

Protein methylation is receiving more and more attention for its important regulating role in diverse biological processes including epigenetic regulation of gene transcription, RNA processing, DNA damage repair, and signal transduction. Global analysis of protein methylation at the proteome level requires the enrichment of methylated peptides with various forms; unfortunately, the immunoaffinity purification method can only enrich a subset of them due to lacking of pan specific antibody. Because methylation does not significantly alter the physicochemical properties of arginine or lysine residues, chemical approach for global methylome analysis is still at infancy. In this study, by exploiting the fact that the methylation on Arg and Lys prohibiting the cleavage by proteases for these sites, we developed an antibody-free method to enrich methylated peptides, which enabled the identification of 887 methylation forms on 768 sites from HepG2 cells. This technique allows the simultaneous analysis of both Lys and Arg methylation while it has better performance for the identification of Arg methylation. It should find broad applications in studying methylation regulated biological processes.

中文翻译：

蛋白质甲基化全局分析的无抗体方法

蛋白质甲基化由于其在多种生物过程中的重要调控作用而受到越来越多的关注，这些过程包括基因转录的表观遗传调控，RNA加工，DNA损伤修复和信号转导。对蛋白质组水平的蛋白质甲基化进行全局分析需要富集各种形式的甲基化肽。不幸的是，由于缺乏泛特异性抗体，免疫亲和纯化方法只能富集其中的一部分。由于甲基化不会显着改变精氨酸或赖氨酸残基的理化性质，因此用于整体甲基化组分析的化学方法仍处于起步阶段。在这项研究中，通过利用Arg和Lys的甲基化阻止蛋白酶切割这些位点这一事实，我们开发了一种无抗体的方法来富集甲基化的肽，从而能够鉴定来自HepG2细胞的768个位点上的887个甲基化形式。该技术允许同时分析Lys和Arg甲基化，同时具有更好的Arg甲基化识别性能。它应该在研究甲基化调控的生物过程中找到广泛的应用。

更新日期：2016-11-14

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