TY - DATA T1 - Quantitative Phosphoproteomic Analysis of the Tumor Necrosis Factor Pathway PY - 2006/01/06 AU - Greg T. Cantin AU - John D. Venable AU - Daniel Cociorva AU - John R. Yates UR - https://acs.figshare.com/articles/dataset/Quantitative_Phosphoproteomic_Analysis_of_the_Tumor_Necrosis_Factor_Pathway/3245521 DO - 10.1021/pr050270m.s006 L4 - https://ndownloader.figshare.com/files/5079631 KW - isoelectric KW - 145 phosphorylation sites KW - HeLa cells metabolically KW - TNF pathway KW - LC KW - Tumor Necrosis Factor Pathway Protein phosphorylation KW - PKA KW - proteomic studies KW - metal affinity chromatography KW - phosphorylation events KW - tumor necrosis factor KW - phosphoproteomic analysis KW - Quantitative Phosphoproteomic Analysis KW - IMAC KW - protein samples N2 - 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. Keywords: isoelectric focusing • IMAC • phosphoproteome • metabolic labeling • TNF • PKA • apoptosis ER -