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Quantitative phosphoproteomic analysis of the tumor necrosis factor pathway.

ABSTRACT: Protein phosphorylation has become a focus of many proteomic studies due to the central role that it plays in biology. We combine peptide-based gel-free isoelectric focusing and immobilized metal affinity chromatography to enhance the detection of phosphorylation events within complex protein samples using LC-MS. This method is then used to carry out a quantitative phosphoproteomic analysis of the tumor necrosis factor (TNF) pathway using HeLa cells metabolically labeled with 15N-containing amino acids, where 145 phosphorylation sites were found to be up-regulated upon the activation of the TNF pathway.

SUBMITTER: Cantin GT

PROVIDER: S-EPMC2570265 | biostudies-literature | 2006 Jan

REPOSITORIES: biostudies-literature

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Quantitative phosphoproteomic analysis of the tumor necrosis factor pathway.

Cantin Greg T GT Venable John D JD Cociorva Daniel D Yates John R JR

Journal of proteome research 20060101 1

Protein phosphorylation has become a focus of many proteomic studies due to the central role that it plays in biology. We combine peptide-based gel-free isoelectric focusing and immobilized metal affinity chromatography to enhance the detection of phosphorylation events within complex protein samples using LC-MS. This method is then used to carry out a quantitative phosphoproteomic analysis of the tumor necrosis factor (TNF) pathway using HeLa cells metabolically labeled with 15N-containing amin ...[more]

PMID: 16396503

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