Combining protein-based IMAC, peptide-based IMAC, and MudPIT for efficient phosphoproteomic analysis

被引：128

作者：

Cantin, Greg T. ^{[1
]}

Yi, Wei ^{[2
]}

Lu, Bingwen ^{[1
]}

Park, Sung Kyu ^{[1
]}

Xu, Tao ^{[1
]}

Lee, Jiing-Dwan ^{[2
]}

Yates, John R., III ^{[1
]}

机构：

[1] Scripps Res Inst, Dept Cell Biol, La Jolla, CA 92037 USA

[2] Scripps Res Inst, Dept Immunol, La Jolla, CA 92037 USA

来源：

JOURNAL OF PROTEOME RESEARCH | 2008年 / 7卷 / 03期

关键词：

phosphoproteome; IMAC; SILAC; EGF;

D O I：

10.1021/pr0705441

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Immobilized metal affinity chromatography (IMAC) is a common strategy used for the enrichment of phosphopeptides from digested protein mixtures. However, this strategy by itself is inefficient when analyzing complex protein mixtures. Here, we assess the effectiveness of using protein-based IMAC as a pre-enrichment step prior to peptide-based IMAC. Ultimately, we couple the two IMAC-based enrichments and MudPIT in a quantitative phosphoproteomic analysis of the epidermal growth factor pathway in mammalian cells identifying 4470 unique phosphopeptides containing 4729 phosphorylation sites.

引用

页码：1346 / 1351

页数：6

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