10.1021/pr050270m.s006
Greg T. Cantin
Greg T.
Cantin
John D. Venable
John D.
Venable
Daniel Cociorva
Daniel
Cociorva
John R. Yates
John R.
Yates
Quantitative Phosphoproteomic Analysis of the Tumor Necrosis
Factor Pathway
American Chemical Society
2006
isoelectric
145 phosphorylation sites
HeLa cells metabolically
TNF pathway
LC
Tumor Necrosis Factor Pathway Protein phosphorylation
PKA
proteomic studies
metal affinity chromatography
phosphorylation events
tumor necrosis factor
phosphoproteomic analysis
Quantitative Phosphoproteomic Analysis
IMAC
protein samples
2006-01-06 00:00:00
Dataset
https://acs.figshare.com/articles/dataset/Quantitative_Phosphoproteomic_Analysis_of_the_Tumor_Necrosis_Factor_Pathway/3245521
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 <sup>15</sup>N-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