title: Optimized chemical proteomics assay for kinase inhibitor profiling.
abstract: Solid supported probes have proven to be an efficient tool for chemical proteomics. The kinobeads technology features kinase inhibitors covalently attached to Sepharose for affinity enrichment of kinomes from cell or tissue lysates. This technology, combined with quantitative mass spectrometry, is of particular interest for the profiling of kinase inhibitors. It often leads to the identification of new targets for medicinal chemistry campaigns where it allows a two-in-one binding and selectivity assay. The assay can also uncover resistance mechanisms and molecular sources of toxicity. Here we report on the optimization of the kinobead assay resulting in the combination of five chemical probes and four cell lines to cover half the human kinome in a single assay (∼ 260 kinases). We show the utility and large-scale applicability of the new version of kinobeads by reprofiling the small molecule kinase inhibitors Alvocidib, Crizotinib, Dasatinib, Fasudil, Hydroxyfasudil, Nilotinib, Ibrutinib, Imatinib, and Sunitinib.
authors: Chair of Proteomics and Bioanalytics, Technische Universität München , Freising, Germany.
authors: Guillaume
authors: G
authors: Médard
authors: Fiona
authors: F
authors: Pachl
authors: Benjamin
authors: B
authors: Ruprecht
authors: Susan
authors: S
authors: Klaeger
authors: Stephanie
authors: S
authors: Heinzlmeir
authors: Dominic
authors: D
authors: Helm
authors: Huichao
authors: H
authors: Qiao
authors: Xin
authors: X
authors: Ku
authors: Mathias
authors: M
authors: Wilhelm
authors: Thomas
authors: T
authors: Kuehne
authors: Zhixiang
authors: Z
authors: Wu
authors: Antje
authors: A
authors: Dittmann
authors: Carsten
authors: C
authors: Hopf
authors: Karl
authors: K
authors: Kramer
authors: Bernhard
authors: B
authors: Kuster
mesh_terms: D013058:Mass Spectrometry; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D040901:Proteomics
chemical_list: D047428:Protein Kinase Inhibitors
keywords: affinity matrix; chemical proteomics; drug profiling; kinase inhibitors; kinobeads