title: Activation state-selective kinase inhibitor assay based on ion mobility-mass spectrometry.
abstract: The discovery of activation state dependent kinase inhibitors, which bind specifically to the inactive conformation of the protein, is considered to be a promising pathway to improved cancer treatments. Identifying such inhibitors is challenging, however, because they can have Kd values similar to molecules known to inhibit kinase function by interacting with the active form. Further, while inhibitor induced changes within the kinase tertiary structure are significant, few technologies are able to correctly assign inhibitor binding modes in a high-throughput fashion based exclusively on protein-inhibitor complex formation and changes in local protein structure. We have developed a new assay, using ion mobility-mass spectrometry, capable of both rapidly detecting inhibitor binding and classifying the resultant kinase binding modes. Here, we demonstrate the ability of our approach to classify a broad set of kinase inhibitors, using micrograms of protein, without the need for protein modification or tagging.
authors.affiliation: Department of Chemistry, University of Michigan, Ann Arbor, Michigan 48109, United States.
authors.forename: Jessica N
authors.initials: JN
authors.lastname: Rabuck
authors.forename: Suk-Joon
authors.initials: SJ
authors.lastname: Hyung
authors.forename: Kristin S
authors.initials: KS
authors.lastname: Ko
authors.forename: Christel C
authors.initials: CC
authors.lastname: Fox
authors.forename: Matthew B
authors.initials: MB
authors.lastname: Soellner
authors.forename: Brandon T
authors.initials: BT
authors.lastname: Ruotolo
mesh_terms: D055808:Drug Discovery / Q000379:methods*; D004789:Enzyme Activation; D013058:Mass Spectrometry; D008958:Models, Molecular; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D017434:Protein Structure, Tertiary; D058767:Protein Unfolding; D016315:Proto-Oncogene Proteins c-abl / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism
chemical_list: D047428:Protein Kinase Inhibitors; D016315:Proto-Oncogene Proteins c-abl