title: Structural determinants of substrate selection by the human insulin-receptor protein-tyrosine kinase.
abstract: Using NMR spectroscopy to visualise tyrosine phosphorylation kinetics in real time, we have investigated the sequence-dependent determinants of the selectivity of the human insulin receptor protein-tyrosine kinase for different tyrosine residues. The peptides used encompass the multiple-tyrosine-containing autophosphorylation site sequences from the insulin receptor kinase core domain (Tyr1158, Tyr1162 and Tyr1163) and from its specific C-terminal tail domain (Tyr1328 and Tyr1334). Comparison of the phosphorylation kinetics with those found for the tyrosine residues on a peptide comprising the regulatory tyrosine phosphorylation site of cdc2 points to the role of the primary sequence context of the phosphate acceptor. The particularly deleterious influence of a basic residue immediately C-terminal to the tyrosine is discussed in relation to the autophosphorylation properties of the regulatory loop regions of the insulin and epidermal growth factor receptor kinases. The data further suggest that receptor tyrosine kinase active sites and their substrate targets act in concert to ensure that specific downstream effects are activated.
authors.affiliation: School of Biochemistry, University of Birmingham, England.
authors.forename: N E
authors.initials: NE
authors.lastname: Keane
authors.forename: A
authors.initials: A
authors.lastname: Chavanieu
authors.forename: P G
authors.initials: PG
authors.lastname: Quirk
authors.forename: J S
authors.initials: JS
authors.lastname: Evans
authors.forename: B A
authors.initials: BA
authors.lastname: Levine
authors.forename: B
authors.initials: B
authors.lastname: Calas
authors.forename: L
authors.initials: L
authors.lastname: Wei
authors.forename: L
authors.initials: L
authors.lastname: Ellis
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D004563:Electrochemistry; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019000:Phosphotyrosine; D011487:Protein Conformation; D011972:Receptor, Insulin / Q000737:chemistry* / Q000378:metabolism; D013379:Substrate Specificity; D014443:Tyrosine / Q000031:analogs & derivatives / Q000378:metabolism
chemical_list: D019000:Phosphotyrosine; D014443:Tyrosine; D011972:Receptor, Insulin