Journal
JOURNAL OF PROTEOME RESEARCH
Volume 7, Issue 3, Pages 1346-1351Publisher
AMER CHEMICAL SOC
DOI: 10.1021/pr0705441
Keywords
phosphoproteome; IMAC; SILAC; EGF
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Funding
- NCRR NIH HHS [P41 RR11823] Funding Source: Medline
- NIMH NIH HHS [5R01 MH067880] Funding Source: Medline
- PHS HHS [U0M/DMID-BAA-03] Funding Source: Medline
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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.
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