title: Identification of Kinase-substrate Pairs Using High Throughput Screening.
abstract: We have developed a screening platform to identify dedicated human protein kinases for phosphorylated substrates which can be used to elucidate novel signal transduction pathways. Our approach features the use of a library of purified GST-tagged human protein kinases and a recombinant protein substrate of interest. We have used this technology to identify MAP/microtubule affinity-regulating kinase 2 (MARK2) as the kinase for a glucose-regulated site on CREB-Regulated Transcriptional Coactivator 2 (CRTC2), a protein required for beta cell proliferation, as well as the Axl family of tyrosine kinases as regulators of cell metastasis by phosphorylation of the adaptor protein ELMO. We describe this technology and discuss how it can help to establish a comprehensive map of how cells respond to environmental stimuli.
authors.affiliation: Children's Hospital of Eastern Ontario Research Institute.
authors.forename: Courtney
authors.initials: C
authors.lastname: Reeks
authors.affiliation: Sunnybrook Research Institute, University of Toronto; Department of Biochemistry, University of Toronto; robert.screaton@sri.utoronto.ca.
authors.forename: Robert A
authors.initials: RA
authors.lastname: Screaton
mesh_terms: D005982:Glutathione Transferase / Q000737:chemistry / Q000378:metabolism; D057809:HEK293 Cells; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry / Q000378:metabolism*; D015398:Signal Transduction; D014157:Transcription Factors / Q000737:chemistry / Q000378:metabolism*
chemical_list: C493245:CRTC2 protein, human; D014157:Transcription Factors; D005982:Glutathione Transferase; C094670:MARK2 protein, human; D017346:Protein Serine-Threonine Kinases