title: Characterization of irreversible kinase inhibitors by directly detecting covalent bond formation: a tool for dissecting kinase drug resistance.
abstract: Targeting protein kinases in cancer therapy with irreversible small-molecule inhibitors is moving to the forefront of kinase-inhibitor research and is thought to be an effective means of overcoming mutation-associated drug resistance in epidermal growth factor receptor kinase (EGFR). We generated a detection technique that allows direct measurements of covalent bond formation without relying on kinase activity, thereby allowing the straightforward investigation of the influence of steric clashes on covalent inhibitors in different resistant kinase mutants. The obtained results are discussed together with structural biology and biochemical studies of catalytic activity in both wild-type and gatekeeper mutated kinase variants to draw conclusions about the impact of steric hindrance and increased catalytic activity in drug-resistant kinase variants.
authors.affiliation: Chemical Genomics Centre of the Max Planck Society, Otto-Hahn-Strasse 15, 44227 Dortmund, Germany.
authors.forename: Sabine
authors.initials: S
authors.lastname: Klüter
authors.forename: Jeffrey R
authors.initials: JR
authors.lastname: Simard
authors.forename: Haridas B
authors.initials: HB
authors.lastname: Rode
authors.forename: Christian
authors.initials: C
authors.lastname: Grütter
authors.forename: Vijaykumar
authors.initials: V
authors.lastname: Pawar
authors.forename: Hans C A
authors.initials: HC
authors.lastname: Raaijmakers
authors.forename: Tjeerd A
authors.initials: TA
authors.lastname: Barf
authors.forename: Matthias
authors.initials: M
authors.lastname: Rabiller
authors.forename: Willem A L
authors.initials: WA
authors.lastname: van Otterlo
authors.forename: Daniel
authors.initials: D
authors.lastname: Rauh
mesh_terms: D000818:Animals; D002645:Chickens; D018360:Crystallography, X-Ray; D019008:Drug Resistance, Neoplasm; D066246:ErbB Receptors / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008958:Models, Molecular; D015394:Molecular Structure; D009154:Mutation; D009369:Neoplasms / Q000188:drug therapy; D047428:Protein Kinase Inhibitors / Q000737:chemistry* / Q000494:pharmacology*; D011494:Protein Kinases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D013050:Spectrometry, Fluorescence / Q000379:methods*; D019061:src-Family Kinases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
chemical_list: D047428:Protein Kinase Inhibitors; D011494:Protein Kinases; D066246:ErbB Receptors; D019061:src-Family Kinases