title: Direct binding assay for the detection of type IV allosteric inhibitors of Abl.
abstract: Abelson (Abl) tyrosine kinase is an important cellular enzyme that is rendered constitutively active in the breakpoint cluster region (BCR)-Abl fusion protein, contributing to several forms of leukemia. Although inhibiting BCR-Abl activity with imatinib shows great clinical success, many patients acquire secondary mutations that result in resistance to imatinib. Second-generation inhibitors such as dasatinib and nilotinib can overcome the majority of these mutations but fail to treat patients with an especially prevalent T315I mutation at the gatekeeper position of the kinase domain. However, a combination of nilotinib with an allosteric type IV inhibitor was recently shown to overcome this clinically relevant point mutation. In this study, we present the development of a direct binding assay that enables the straightforward detection of allosteric inhibitors which bind within the myristate pocket of Abl. The assay is amenable to high-throughput screening and exclusively detects the binding of ligands to this unique allosteric site.
authors.affiliation: Chemical Genomics Centre of the Max-Planck-Society, Dortmund, Germany.
authors.forename: Ralf
authors.initials: R
authors.lastname: Schneider
authors.forename: Christian
authors.initials: C
authors.lastname: Becker
authors.forename: Jeffrey R
authors.initials: JR
authors.lastname: Simard
authors.forename: Matthäus
authors.initials: M
authors.lastname: Getlik
authors.forename: Nina
authors.initials: N
authors.lastname: Bohlke
authors.forename: Petra
authors.initials: P
authors.lastname: Janning
authors.forename: Daniel
authors.initials: D
authors.lastname: Rauh
mesh_terms: D008958:Models, Molecular; D015394:Molecular Structure; D047428:Protein Kinase Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D016315:Proto-Oncogene Proteins c-abl / Q000037:antagonists & inhibitors* / Q000378:metabolism; D013329:Structure-Activity Relationship
chemical_list: D047428:Protein Kinase Inhibitors; D016315:Proto-Oncogene Proteins c-abl