title: Characterization of a conserved structural determinant controlling protein kinase sensitivity to selective inhibitors.
abstract: Some protein kinases are known to acquire resistance to selective small molecule inhibitors upon mutation of a conserved threonine at the ATP binding site to a larger residue. Here, we performed a comprehensive mutational analysis of this structural element and determined the cellular sensitivities of several disease-relevant tyrosine kinases against various inhibitors. Mutant kinases possessing a larger side chain at the critical site showed resistance to most compounds tested, such as ZD1839, PP1, AG1296, STI571, and a pyrido[2,3-d]pyrimidine inhibitor. In contrast, indolinones affected both wild-type and mutant kinases with similar potencies. Resistant mutants were established for pharmacological analysis of betaPDGF receptor-mediated signaling and allowed the generation of a drug-inducible system of cellular Src kinase activity. Our data establish a conserved structural determinant of protein kinase sensitivity relevant for both signal transduction research and drug development.
authors.affiliation: Axxima Pharmaceuticals AG, Max-Lebsche-Platz 32, 81377 Munich, Germany.
authors.forename: Stephanie
authors.initials: S
authors.lastname: Blencke
authors.forename: Birgit
authors.initials: B
authors.lastname: Zech
authors.forename: Ola
authors.initials: O
authors.lastname: Engkvist
authors.forename: Zoltán
authors.initials: Z
authors.lastname: Greff
authors.forename: László
authors.initials: L
authors.lastname: Orfi
authors.forename: Zoltán
authors.initials: Z
authors.lastname: Horváth
authors.forename: György
authors.initials: G
authors.lastname: Kéri
authors.forename: Axel
authors.initials: A
authors.lastname: Ullrich
authors.forename: Henrik
authors.initials: H
authors.lastname: Daub
mesh_terms: D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D017124:Conserved Sequence / Q000235:genetics / Q000502:physiology; D004815:Epidermal Growth Factor / Q000235:genetics; D008969:Molecular Sequence Data; D047428:Protein Kinase Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors* / Q000737:chemistry* / Q000235:genetics; D020794:Receptor Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics; D051496:Receptor, Fibroblast Growth Factor, Type 1; D020797:Receptor, Platelet-Derived Growth Factor beta / Q000235:genetics; D017468:Receptors, Fibroblast Growth Factor / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics; D016415:Sequence Alignment; D015398:Signal Transduction; D019061:src-Family Kinases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics
chemical_list: D047428:Protein Kinase Inhibitors; D017468:Receptors, Fibroblast Growth Factor; D004815:Epidermal Growth Factor; D011505:Protein-Tyrosine Kinases; D020794:Receptor Protein-Tyrosine Kinases; D051496:Receptor, Fibroblast Growth Factor, Type 1; D020797:Receptor, Platelet-Derived Growth Factor beta; D019061:src-Family Kinases