Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
4
pubmed:dateCreated
2001-4-3
pubmed:abstractText
Reversible protein phosphorylation has been known for some time to control a wide range of biological functions and activities. Thus determination of the site(s) of protein phosphorylation has been an essential step in the analysis of the control of many biological systems. However, direct determination of individual phosphorylation sites occurring on phosphoproteins in vivo has been difficult to date, typically requiring the purification to homogeneity of the phosphoprotein of interest before analysis. Thus, there has been a substantial need for a more rapid and general method for the analysis of protein phosphorylation in complex protein mixtures. Here we describe such an approach to protein phosphorylation analysis. It consists of three steps: (1) selective phosphopeptide isolation from a peptide mixture via a sequence of chemical reactions, (2) phosphopeptide analysis by automated liquid chromatography-tandem mass spectrometry (LC-MS/MS), and (3) identification of the phosphoprotein and the phosphorylated residue(s) by correlation of tandem mass spectrometric data with sequence databases. By utilizing various phosphoprotein standards and a whole yeast cell lysate, we demonstrate that the method is equally applicable to serine-, threonine- and tyrosine-phosphorylated proteins, and is capable of selectively isolating and identifying phosphopeptides present in a highly complex peptide mixture.
pubmed:grant
pubmed:commentsCorrections
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Apr
pubmed:issn
1087-0156
pubmed:author
pubmed:issnType
Print
pubmed:volume
19
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
375-8
pubmed:dateRevised
2007-11-14
pubmed:meshHeading
pubmed-meshheading:11283598-Amino Acid Sequence, pubmed-meshheading:11283598-Biochemistry, pubmed-meshheading:11283598-Caseins, pubmed-meshheading:11283598-Databases, Factual, pubmed-meshheading:11283598-Fungal Proteins, pubmed-meshheading:11283598-Gas Chromatography-Mass Spectrometry, pubmed-meshheading:11283598-Glucose, pubmed-meshheading:11283598-Glutathione Transferase, pubmed-meshheading:11283598-Mass Spectrometry, pubmed-meshheading:11283598-Models, Chemical, pubmed-meshheading:11283598-Molecular Sequence Data, pubmed-meshheading:11283598-Peptides, pubmed-meshheading:11283598-Phosphorylation, pubmed-meshheading:11283598-Recombinant Fusion Proteins, pubmed-meshheading:11283598-Saccharomyces cerevisiae, pubmed-meshheading:11283598-Serine, pubmed-meshheading:11283598-Threonine, pubmed-meshheading:11283598-Time Factors, pubmed-meshheading:11283598-Tyrosine
pubmed:year
2001
pubmed:articleTitle
A systematic approach to the analysis of protein phosphorylation.
pubmed:affiliation
Department of Molecular Biotechnology, University of Washington, Seattle, WA 98195, USA.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, U.S. Gov't, Non-P.H.S., Research Support, Non-U.S. Gov't