4.7 Article

Antibody-free enrichment method for proteome-wide analysis of endogenous SUMOylation sites

期刊

ANALYTICA CHIMICA ACTA
卷 1154, 期 -, 页码 -

出版社

ELSEVIER
DOI: 10.1016/j.aca.2021.338324

关键词

Endogenous SUMOylation; Antibody-free enrichment; Strong anion exchange; Comprehensive profiling

资金

  1. National Key R&D Program of China [2017YFA0505003]
  2. National Natural Science Foundation of China [21675153, 91753110, 21725506]
  3. Innovation Academy for Precision Measurement Science and Technology, CAS

向作者/读者索取更多资源

The study introduces an antibody-free strategy to simultaneously enrich and profile endogenous SUMO1/2/3-modified peptides, allowing for a comprehensive mapping of various endogenous SUMOylation sites.
SUMOylation is a reversible post-translational modification that plays crucial roles in numerous cellular processes. Although anti-SUMO antibodies have been applied to analyze exogenous and endogenous SUMOylation, such immunoprecipitation enrichment strategy is applicable only for the enrichment of one specific SUMO type in mammalian cells, unable to map the global landscape of all endogenous SUMOylation simultaneously. To address this issue, we proposed an antibody-free strategy to enrich and profile endogenous SUMO1/2/3-modified peptides simultaneously. Upon trypsin digestion, the SUMO1- and SUMO2/3-modified peptides contained SUMO remnants with 7 and 9 acidic amino acids respectively, which carried more negative charges at high pH and could interact with strong anion exchange (SAX) materials more strongly than non-SUMOylated peptides, thus enabling the specific enrichment of endogenous SUMOylated peptides. Followed by the secondary digestion with Asp-N/Glu-C to generate smaller SUMOylated peptides with proper length for MS identification, off-line high-pH C18 prefractionation and low pH nanoRPLC-ESI-MS/MS analysis, 177 SUMO1-modified sites and 74 SUMO2/3-modified sites were unbiasedly identified in HeLa cell lysate. To the best of our knowledge, this was the first antibody-free strategy to comprehensively profile various endogenous SUMOylation sites, demonstrating the great potential in the comprehensive analysis of endogenous SUMOylation across various species and organs, which might further facilitate the understanding of SUMO's function in physiology and pathology. (C) 2021 Elsevier B.V. All rights reserved.

作者

我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。

评论

主要评分

4.7
评分不足

次要评分

新颖性
-
重要性
-
科学严谨性
-
评价这篇论文

推荐

暂无数据
暂无数据