Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
21
pubmed:dateCreated
2008-11-4
pubmed:abstractText
The liver is a central organ involved in many aspects of physiology and disease. Signaling properties of hepatocytes, the main liver cell type, are of special interest in metabolic diseases and in regeneration. For this reason we investigated the phosphoproteome of the mouse liver cell line Hepa1-6 by stable isotope labeling by amino acids in cell culture (SILAC) and high resolution MS. Using stringent statistical evaluation criteria, we obtained 5433 phosphorylation sites on 1808 proteins. The phosphoproteome encompasses all major protein classes, including a large number of transcription factors. We compared control and phosphatase inhibitor treated cells by SILAC. This enabled ready identification of in vivo phosphorylation sites by sequencing the more abundant, inhibitor induced version of the peptide while still observing the endogenous version. We employed a mixture of pervanadate for blocking protein tyrosine phosphatases (PTPs) and calyculin A and deltamethrin for blocking the activities of serine/threonine phosphatases. Interestingly, these commonly used inhibitors in standard concentrations affected only 28% of the phosphopeptides by at least two-fold. The unaffected sites may be substrates of phosphatases that are not efficiently inhibited, have slow kinetic or sites that are almost stoichiometric in normally growing cells. Finally, we devised a triple labeling strategy comprising control cells, stimulated cells, and phosphatase treated cells to derive an upper bound on phosphorylation occupancy.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
http://linkedlifedata.com/resource/pubmed/chemical/Insulin, http://linkedlifedata.com/resource/pubmed/chemical/Nitriles, http://linkedlifedata.com/resource/pubmed/chemical/Oxazoles, http://linkedlifedata.com/resource/pubmed/chemical/Phosphoprotein Phosphatases, http://linkedlifedata.com/resource/pubmed/chemical/Phosphoproteins, http://linkedlifedata.com/resource/pubmed/chemical/Protein Tyrosine Phosphatases, http://linkedlifedata.com/resource/pubmed/chemical/Proteome, http://linkedlifedata.com/resource/pubmed/chemical/Pyrethrins, http://linkedlifedata.com/resource/pubmed/chemical/Transcription Factors, http://linkedlifedata.com/resource/pubmed/chemical/Vanadates, http://linkedlifedata.com/resource/pubmed/chemical/calyculin A, http://linkedlifedata.com/resource/pubmed/chemical/decamethrin, http://linkedlifedata.com/resource/pubmed/chemical/pervanadate
pubmed:status
MEDLINE
pubmed:month
Nov
pubmed:issn
1615-9861
pubmed:author
pubmed:issnType
Electronic
pubmed:volume
8
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
4534-46
pubmed:dateRevised
2011-11-17
pubmed:meshHeading
pubmed-meshheading:18846507-Animals, pubmed-meshheading:18846507-Cell Line, Tumor, pubmed-meshheading:18846507-Chromatography, Liquid, pubmed-meshheading:18846507-Computational Biology, pubmed-meshheading:18846507-Hepatocytes, pubmed-meshheading:18846507-Insulin, pubmed-meshheading:18846507-Liver, pubmed-meshheading:18846507-Mice, pubmed-meshheading:18846507-Nitriles, pubmed-meshheading:18846507-Oxazoles, pubmed-meshheading:18846507-Phosphoprotein Phosphatases, pubmed-meshheading:18846507-Phosphoproteins, pubmed-meshheading:18846507-Phosphorylation, pubmed-meshheading:18846507-Protein Tyrosine Phosphatases, pubmed-meshheading:18846507-Proteome, pubmed-meshheading:18846507-Pyrethrins, pubmed-meshheading:18846507-Substrate Specificity, pubmed-meshheading:18846507-Tandem Mass Spectrometry, pubmed-meshheading:18846507-Transcription Factors, pubmed-meshheading:18846507-Vanadates
pubmed:year
2008
pubmed:articleTitle
Quantitative phosphoproteome analysis of a mouse liver cell line reveals specificity of phosphatase inhibitors.
pubmed:affiliation
Department of Proteomics and Signal Transduction, Max-Planck Institute for Biochemistry, Martinsried, Germany.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't