UbiSite approach for comprehensive mapping of lysine and N-terminal ubiquitination sites

被引：357

作者：

Akimov, Vyacheslav ^{[1
]}

Barrio-Hernandez, Inigo ^{[1
]}

Hansen, Sten V. F. ^{[1
]}

Hallenborg, Philip ^{[1
]}

Pedersen, Anna-Kathrine ^{[2
]}

Bekker-Jensen, Dorte B. ^{[2
]}

Puglia, Michele ^{[1
]}

Christensen, Stine D. K. ^{[1
]}

Vanselow, Jens T. ^{[1
]}

Nielsen, Mogens M. ^{[1
]}

Kratchmarova, Irina ^{[1
]}

Kelstrup, Christian D. ^{[2
]}

Olsen, Jesper, V ^{[2
]}

Blagoev, Blagoy ^{[1
]}

机构：

[1] Univ Southern Denmark, Dept Biochem & Mol Biol, Odense, Denmark

[2] Univ Copenhagen, Novo Nordisk Fdn Ctr Prot Res, Fac Hlth & Med Sci, Copenhagen, Denmark

来源：

NATURE STRUCTURAL & MOLECULAR BIOLOGY | 2018年 / 25卷 / 07期

基金：

英国医学研究理事会; 新加坡国家研究基金会; 欧盟地平线“2020”;

关键词：

QUANTITATIVE PROTEOMICS REVEALS; PROTEASOME PATHWAY; TAGGED UBIQUITIN; PROTEIN UBIQUITINATION; EXCHANGE SYSTEM; DEGRADATION; QUANTIFICATION; DEUBIQUITINASE; PURIFICATION; KINASE;

D O I：

10.1038/s41594-018-0084-y

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Ubiquitination is a post-translational modification (PTM) that is essential for balancing numerous physiological processes. To enable delineation of protein ubiquitination at a site-specific level, we generated an antibody, denoted UbiSite, recognizing the C-terminal 13 amino acids of ubiquitin, which remain attached to modified peptides after proteolytic digestion with the endoproteinase LysC. Notably, UbiSite is specific to ubiquitin. Furthermore, besides ubiquitination on lysine residues, protein N-terminal ubiquitination is readily detected as well. By combining UbiSite enrichment with sequential LysC and trypsin digestion and high-accuracy MS, we identified over 63,000 unique ubiquitination sites on 9,200 proteins in two human cell lines. In addition to uncovering widespread involvement of this PTM in all cellular aspects, the analyses reveal an inverse association between protein N-terminal ubiquitination and acetylation, as well as a complete lack of correlation between changes in protein abundance and alterations in ubiquitination sites upon proteasome inhibition.

引用

页码：631 / +

页数：13

共 69 条

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