title: Catalytic activity and transformation potential of v-Src require arginine 385 in the substrate binding pocket.
abstract: Tyrosine kinase are important mediators of signal transduction in eukaryotic cells. In order to better understand the mechanism of catalysis we studied a set of mutants of the prototype tyrosine kinase, the c-Src protein, a homologue of the Rous Sarcoma virus oncogene. Based on an X-ray structure of cAMP-dependent protein kinase (cAPK) we mutated an arginine residue conserved in subdomain VI of all known kinases to a non-charged residue. This residue coordinates phosphate of the autophosphorylation site located in subdomain VII of cAPK and this interaction has been proposed to be crucial for substrate binding. The mutant R385A of c-Src had low kinase activity towards exogenous substrates yet was able to autophosphorylate at tyrosine 416. When introduced into an activated v-src gene the R385A mutation totally blocked cell transformation. Our data suggest that the function of the conserved arginine 385 is to coordinate the phosphate of the autophosphorylation site and to provide in this way a stable template for substrate binding.
authors.affiliation: Friedrich Miescher-Institute, Basel, Switzerland.
authors.forename: M
authors.initials: M
authors.lastname: Senften
authors.forename: G
authors.initials: G
authors.lastname: Schenker
authors.forename: J M
authors.initials: JM
authors.lastname: Sowadski
authors.forename: K
authors.initials: K
authors.lastname: Ballmer-Hofer
mesh_terms: D016475:3T3 Cells; D000818:Animals; D001120:Arginine / Q000378:metabolism*; D001358:Avian Sarcoma Viruses / Q000235:genetics / Q000502:physiology; D001483:Base Sequence; D001665:Binding Sites; D002384:Catalysis; D002471:Cell Transformation, Neoplastic; D002472:Cell Transformation, Viral; D002478:Cells, Cultured; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D009838:Oligodeoxyribonucleotides; D015688:Oncogene Protein pp60(v-src) / Q000235:genetics / Q000502:physiology*; D016392:Proto-Oncogene Proteins pp60(c-src) / Q000502:physiology*; D051381:Rats
chemical_list: D009838:Oligodeoxyribonucleotides; D001120:Arginine; D015688:Oncogene Protein pp60(v-src); D016392:Proto-Oncogene Proteins pp60(c-src)