Systematic and Quantitative Assessment of the Ubiquitin-Modified Proteome

被引：1286

作者：

Kim, Woong ^{[1
]}

Bennett, Eric J. ^{[1
,2
]}

Huttlin, Edward L. ^{[1
]}

Guo, Ailan ^{[3
]}

Li, Jing ^{[3
]}

Possemato, Anthony ^{[3
]}

Sowa, Mathew E. ^{[1
,2
]}

Rad, Ramin ^{[1
]}

Rush, John ^{[3
]}

Comb, Michael J. ^{[3
]}

Harper, J. Wade ^{[1
,2
]}

Gygi, Steven P. ^{[2
]}

机构：

[1] Harvard Univ, Sch Med, Dept Cell Biol, Boston, MA 02115 USA

[2] Harvard Univ, Sch Med, Dept Pathol, Boston, MA USA

[3] Cell Signaling Technol, Danvers, MA USA

来源：

MOLECULAR CELL | 2011年 / 44卷 / 02期

关键词：

PHOSPHORYLATION; IDENTIFICATION; PROTEINS; TARGETS; NEDD8; ACETYLATION; REVEALS; CHAINS; CELLS;

D O I：

10.1016/j.molcel.2011.08.025

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：

Despite the diverse biological pathways known to be regulated by ubiquitylation, global identification of substrates that are targeted for ubiquitylation has remained a challenge. To globally characterize the human ubiquitin-modified proteome (ubiquitinome), we utilized a monoclonal antibody that recognizes diglycine (diGly)-containing isopeptides following trypsin digestion. We identify similar to 19,000 diGly-modified lysine residues within similar to 5000 proteins. Using quantitative proteomics we monitored temporal changes in diGly site abundance in response to both proteasomal and translational inhibition, indicating both a dependence on ongoing translation to observe alterations in site abundance and distinct dynamics of individual modified lysines in response to proteasome inhibition. Further, we demonstrate that quantitative diGly proteomics can be utilized to identify substrates for cullin-RING ubiquitin ligases. Interrogation of the ubiquitinome allows for not only a quantitative assessment of alterations in protein homeo-stasis fidelity, but also identification of substrates for individual ubiquitin pathway enzymes.

引用

页码：325 / 340

页数：16

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