title: Identification of tyrosine phosphatases that dephosphorylate the insulin receptor. A brute force approach based on "substrate-trapping" mutants.
abstract: Many pharmacologically important receptors, including all cytokine receptors, signal via tyrosine (auto)phosphorylation, followed by resetting to their original state through the action of protein tyrosine phosphatases (PTPs). Establishing the specificity of PTPs for receptor substrates is critical both for understanding how signaling is regulated and for the development of specific PTP inhibitors that act as ligand mimetics. We have set up a systematic approach for finding PTPs that are specific for a receptor and have validated this approach with the insulin receptor kinase. We have tested nearly all known human PTPs (45) in a membrane binding assay, using "substrate-trapping" PTP mutants. These results, combined with secondary dephosphorylation tests, confirm and extend earlier findings that PTP-1b and T-cell PTP are physiological enzymes for the insulin receptor kinase. We demonstrate that this approach can rapidly reduce the number of PTPs that have a particular receptor or other phosphoprotein as their substrate.
authors.affiliation: Serono Pharmaceutical Research Institute, 14, chemin des Aulx, 1228 Plan-les-Ouates, Geneva 1228, Switzerland.
authors.forename: S
authors.initials: S
authors.lastname: Wälchli
authors.forename: M L
authors.initials: ML
authors.lastname: Curchod
authors.forename: R P
authors.initials: RP
authors.lastname: Gobert
authors.forename: S
authors.initials: S
authors.lastname: Arkinstall
authors.forename: R
authors.initials: R
authors.lastname: Hooft van Huijsduijnen
mesh_terms: D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D003101:Collodion; D018076:DNA, Complementary; D006801:Humans; D008969:Molecular Sequence Data; D016296:Mutagenesis; D010766:Phosphorylation; D017027:Protein Tyrosine Phosphatases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011972:Receptor, Insulin / Q000378:metabolism*; D013379:Substrate Specificity
chemical_list: D018076:DNA, Complementary; D003101:Collodion; D011972:Receptor, Insulin; D017027:Protein Tyrosine Phosphatases