title: Covalent cross-linking of kinases with their corresponding peptide substrates.
abstract: Protein phosphorylation represents the most dominant and evolutionary conserved posttranslational modification for information transfer in cells and organisms. The human genome encodes >500 protein kinases, and thousands of phosphorylation sites are present in mammalian proteome. To develop a global view of phosphorylation network, there is a need to map the connectivity between kinases and phosphoproteome. We developed a chemical kinase-substrate cross-linker 1 that converts transient kinase-substrate interactions into a covalently linked kinase-substrate complex in vitro and in the presence of cell lysates. The method can be applied to identify unknown upstream kinases responsible for phosphorylation events in cell lysates.
authors.affiliation: Department of Chemistry, Northwestern University, Evanston, IL, USA.
authors.forename: Alexander V
authors.initials: AV
authors.lastname: Statsuk
authors.forename: Kevan M
authors.initials: KM
authors.lastname: Shokat
mesh_terms: D045744:Cell Line, Tumor; D003432:Cross-Linking Reagents / Q000138:chemical synthesis / Q000378:metabolism; D003545:Cysteine / Q000378:metabolism; D057075:Enzyme Assays / Q000379:methods*; D005456:Fluorescent Dyes / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D010455:Peptides / Q000138:chemical synthesis / Q000378:metabolism*; D010766:Phosphorylation; D010770:Phosphotransferases / Q000378:metabolism*; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D013194:Staining and Labeling; D013379:Substrate Specificity
chemical_list: D003432:Cross-Linking Reagents; D005456:Fluorescent Dyes; D010455:Peptides; D010770:Phosphotransferases; D051057:Proto-Oncogene Proteins c-akt; D003545:Cysteine