Linked Life Data

19640851

Source:http://linkedlifedata.com/resource/pubmed/id/19640851

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PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
12
pubmed:dateCreated
2009-12-7
pubmed:abstractText
Stimulated by its physiological ligand, hepatocyte growth factor, the transmembrane receptor tyrosine kinase Met activates a signaling machinery that leads to mitogenic, motogenic, and morphogenic responses. Remarkably, the food-borne human pathogen Listeria monocytogenes also promotes autophosphorylation of Met through its virulence factor internalin B (InlB) and subsequently exploits Met signaling to induce phagocytosis into a broad range of host cells. Although the interaction between InlB and Met has been studied in detail, the signaling specificity of components involved in InlB-triggered cellular responses remains poorly characterized. The analysis of regulated phosphorylation events on protein kinases is therefore of particular relevance, although this could not as yet be characterized systematically by proteomics. Here, we implemented a new pyridopyrimidine-based strategy that enabled the efficient capture of a considerable subset of the human kinome in a robust one-step affinity chromatographic procedure. Additionally, and to gain functional insights into the InlB/Met-induced bacterial invasion process, a quantitative survey of the phosphorylation pattern of these protein kinases was accomplished. In total, the experimental design of this study comprises affinity chromatographic procedures for the systematic enrichment of kinases, as well as phosphopeptides; the quantification of all peptides based on the iTRAQ reporter system; and a rational statistical strategy to evaluate the quality of phosphosite regulations. With this improved chemical proteomics strategy, we determined and relatively quantified 143 phosphorylation sites detected on 94 human protein kinases. Interestingly, InlB-mediated signaling shows striking similarities compared with the natural ligand hepatocyte growth factor that was intensively studied in the past. In addition, this systematic approach suggests a new subset of protein kinases including Nek9, which are differentially phosphorylated after short time (4-min) treatment of cells with the Met-activating InlB(321). Thus, this quantitative phosphokinome study suggests a general, hypothesis-free concept for the detection of dynamically regulated protein kinases as novel signaling components involved in host-pathogen interactions.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Dec
pubmed:issn
1535-9484
pubmed:author
pubmed:issnType
Electronic
pubmed:volume
8
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
2778-95
pubmed:dateRevised
2011-3-3
pubmed:meshHeading
pubmed-meshheading:19640851-Amino Acid Sequence, pubmed-meshheading:19640851-Bacterial Proteins, pubmed-meshheading:19640851-Chromatography, Affinity, pubmed-meshheading:19640851-Chromatography, Liquid, pubmed-meshheading:19640851-Cluster Analysis, pubmed-meshheading:19640851-HeLa Cells, pubmed-meshheading:19640851-Host-Pathogen Interactions, pubmed-meshheading:19640851-Humans, pubmed-meshheading:19640851-Isotope Labeling, pubmed-meshheading:19640851-Ligands, pubmed-meshheading:19640851-Listeria monocytogenes, pubmed-meshheading:19640851-Mass Spectrometry, pubmed-meshheading:19640851-Membrane Proteins, pubmed-meshheading:19640851-Molecular Sequence Data, pubmed-meshheading:19640851-Peptides, pubmed-meshheading:19640851-Phosphoproteins, pubmed-meshheading:19640851-Phosphorylation, pubmed-meshheading:19640851-Protein Kinase Inhibitors, pubmed-meshheading:19640851-Proteomics, pubmed-meshheading:19640851-Proto-Oncogene Proteins c-met, pubmed-meshheading:19640851-Signal Transduction
pubmed:year
2009
pubmed:articleTitle
Quantitative phosphokinome analysis of the Met pathway activated by the invasin internalin B from Listeria monocytogenes.
pubmed:affiliation
Department of Cell Biology, Helmholtz Centre for Infection Research, 38124 Braunschweig, Germany.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't