Proteomics analysis of protein kinases by target class-selective prefractionation and tandem mass spectrometry.

2.50
Hdl Handle:
http://hdl.handle.net/10033/19756
Title:
Proteomics analysis of protein kinases by target class-selective prefractionation and tandem mass spectrometry.
Authors:
Wissing, Josef; Jänsch, Lothar ( 0000-0002-5655-1181 ) ; Nimtz, Manfred; Dieterich, Guido; Hornberger, Renate; Kéri, György; Wehland, Jürgen; Daub, Henrik
Abstract:
Protein kinases constitute a large superfamily of enzymes with key regulatory functions in nearly all signal transmission processes of eukaryotic cells. However, due to their relatively low abundance compared with the vast majority of cellular proteins, currently available proteomics techniques do not permit the comprehensive biochemical characterization of protein kinases. To address these limitations, we have developed a prefractionation strategy that uses a combination of immobilized low molecular weight inhibitors for the selective affinity capture of protein kinases. This approach resulted in the direct purification of cell type-specific sets of expressed protein kinases, and more than 140 different members of this enzyme family could be detected by LC-MS/MS. Furthermore the enrichment technique combined with phosphopeptide fractionation led to the identification of more than 200 different phosphorylation sites on protein kinases, which often remain occluded in global phosphoproteome analysis. As the phosphorylation states of protein kinases can provide a readout for the signaling activities within a cellular system, kinase-selective phosphoproteomics based on the procedures described here has the potential to become an important tool in signal transduction analysis.
Affiliation:
Department of Cell Biology, Helmholtz Centre for Infection Research (HZI), Inhoffenstrasse 7, 38124 Braunschweig, Germany.
Citation:
Proteomics analysis of protein kinases by target class-selective prefractionation and tandem mass spectrometry. 2007, 6 (3):537-47 Mol. Cell Proteomics
Journal:
Molecular & cellular proteomics : MCP
Issue Date:
Mar-2007
URI:
http://hdl.handle.net/10033/19756
DOI:
10.1074/mcp.T600062-MCP200
PubMed ID:
17192257
Type:
Article
Language:
en
ISSN:
1535-9476
Appears in Collections:
publications of the research group cellular proteom research (CPRO)

Full metadata record

DC FieldValue Language
dc.contributor.authorWissing, Josefen
dc.contributor.authorJänsch, Lotharen
dc.contributor.authorNimtz, Manfreden
dc.contributor.authorDieterich, Guidoen
dc.contributor.authorHornberger, Renateen
dc.contributor.authorKéri, Györgyen
dc.contributor.authorWehland, Jürgenen
dc.contributor.authorDaub, Henriken
dc.date.accessioned2008-03-05T09:52:10Zen
dc.date.available2008-03-05T09:52:10Zen
dc.date.issued2007-03en
dc.identifier.citationProteomics analysis of protein kinases by target class-selective prefractionation and tandem mass spectrometry. 2007, 6 (3):537-47 Mol. Cell Proteomicsen
dc.identifier.issn1535-9476en
dc.identifier.pmid17192257en
dc.identifier.doi10.1074/mcp.T600062-MCP200en
dc.identifier.urihttp://hdl.handle.net/10033/19756en
dc.description.abstractProtein kinases constitute a large superfamily of enzymes with key regulatory functions in nearly all signal transmission processes of eukaryotic cells. However, due to their relatively low abundance compared with the vast majority of cellular proteins, currently available proteomics techniques do not permit the comprehensive biochemical characterization of protein kinases. To address these limitations, we have developed a prefractionation strategy that uses a combination of immobilized low molecular weight inhibitors for the selective affinity capture of protein kinases. This approach resulted in the direct purification of cell type-specific sets of expressed protein kinases, and more than 140 different members of this enzyme family could be detected by LC-MS/MS. Furthermore the enrichment technique combined with phosphopeptide fractionation led to the identification of more than 200 different phosphorylation sites on protein kinases, which often remain occluded in global phosphoproteome analysis. As the phosphorylation states of protein kinases can provide a readout for the signaling activities within a cellular system, kinase-selective phosphoproteomics based on the procedures described here has the potential to become an important tool in signal transduction analysis.en
dc.language.isoenen
dc.subject.meshCell Line, Tumoren
dc.subject.meshChromatography, Liquiden
dc.subject.meshHumansen
dc.subject.meshPhosphopeptidesen
dc.subject.meshPhosphorylationen
dc.subject.meshProtein Kinase Inhibitorsen
dc.subject.meshProtein Kinasesen
dc.subject.meshProteomicsen
dc.subject.meshTandem Mass Spectrometryen
dc.titleProteomics analysis of protein kinases by target class-selective prefractionation and tandem mass spectrometry.en
dc.typeArticleen
dc.contributor.departmentDepartment of Cell Biology, Helmholtz Centre for Infection Research (HZI), Inhoffenstrasse 7, 38124 Braunschweig, Germany.en
dc.identifier.journalMolecular & cellular proteomics : MCPen

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