title: Chemical Proteomics and Structural Biology Define EPHA2 Inhibition by Clinical Kinase Drugs.
abstract: The receptor tyrosine kinase EPHA2 (Ephrin type-A receptor 2) plays important roles in oncogenesis, metastasis, and treatment resistance, yet therapeutic targeting, drug discovery, or investigation of EPHA2 biology is hampered by the lack of appropriate inhibitors and structural information. Here, we used chemical proteomics to survey 235 clinical kinase inhibitors for their kinase selectivity and identified 24 drugs with submicromolar affinities for EPHA2. NMR-based conformational dynamics together with nine new cocrystal structures delineated drug-EPHA2 interactions in full detail. The combination of selectivity profiling, structure determination, and kinome wide sequence alignment allowed the development of a classification system in which amino acids in the drug binding site of EPHA2 are categorized into key, scaffold, potency, and selectivity residues. This scheme should be generally applicable in kinase drug discovery, and we anticipate that the provided information will greatly facilitate the development of selective EPHA2 inhibitors in particular and the repurposing of clinical kinase inhibitors in general.
authors: Chair of Proteomics and Bioanalytics, Technical University of Munich , 85354 Freising, Germany.|German Cancer Consortium (DKTK) , 69120 Heidelberg, Germany.|German Cancer Research Center (DKFZ) , 69120 Heidelberg, Germany.
authors: Stephanie
authors: S
authors: Heinzlmeir
authors: Center for Biomolecular Magnetic Resonance, Johann Wolfgang Goethe University , 60438 Frankfurt, Germany.|German Cancer Consortium (DKTK) , 69120 Heidelberg, Germany.|German Cancer Research Center (DKFZ) , 69120 Heidelberg, Germany.
authors: Denis
authors: D
authors: Kudlinzki
authors: Center for Biomolecular Magnetic Resonance, Johann Wolfgang Goethe University , 60438 Frankfurt, Germany.
authors: Sridhar
authors: S
authors: Sreeramulu
authors: Chair of Proteomics and Bioanalytics, Technical University of Munich , 85354 Freising, Germany.|German Cancer Consortium (DKTK) , 69120 Heidelberg, Germany.|German Cancer Research Center (DKFZ) , 69120 Heidelberg, Germany.
authors: Susan
authors: S
authors: Klaeger
authors: Center for Biomolecular Magnetic Resonance, Johann Wolfgang Goethe University , 60438 Frankfurt, Germany.|German Cancer Consortium (DKTK) , 69120 Heidelberg, Germany.|German Cancer Research Center (DKFZ) , 69120 Heidelberg, Germany.
authors: Santosh Lakshmi
authors: SL
authors: Gande
authors: Center for Biomolecular Magnetic Resonance, Johann Wolfgang Goethe University , 60438 Frankfurt, Germany.
authors: Verena
authors: V
authors: Linhard
authors: Chair of Proteomics and Bioanalytics, Technical University of Munich , 85354 Freising, Germany.
authors: Mathias
authors: M
authors: Wilhelm
authors: Chair of Proteomics and Bioanalytics, Technical University of Munich , 85354 Freising, Germany.
authors: Huichao
authors: H
authors: Qiao
authors: Chair of Proteomics and Bioanalytics, Technical University of Munich , 85354 Freising, Germany.
authors: Dominic
authors: D
authors: Helm
authors: Chair of Proteomics and Bioanalytics, Technical University of Munich , 85354 Freising, Germany.
authors: Benjamin
authors: B
authors: Ruprecht
authors: Center for Biomolecular Magnetic Resonance, Johann Wolfgang Goethe University , 60438 Frankfurt, Germany.|German Cancer Consortium (DKTK) , 69120 Heidelberg, Germany.|German Cancer Research Center (DKFZ) , 69120 Heidelberg, Germany.
authors: Krishna
authors: K
authors: Saxena
authors: Chair of Proteomics and Bioanalytics, Technical University of Munich , 85354 Freising, Germany.
authors: Guillaume
authors: G
authors: Médard
authors: Center for Biomolecular Magnetic Resonance, Johann Wolfgang Goethe University , 60438 Frankfurt, Germany.|German Cancer Consortium (DKTK) , 69120 Heidelberg, Germany.|German Cancer Research Center (DKFZ) , 69120 Heidelberg, Germany.
authors: Harald
authors: H
authors: Schwalbe
authors: Chair of Proteomics and Bioanalytics, Technical University of Munich , 85354 Freising, Germany.|German Cancer Consortium (DKTK) , 69120 Heidelberg, Germany.|German Cancer Research Center (DKFZ) , 69120 Heidelberg, Germany.|Center for Integrated Protein Science Munich (CIPSM) , 85354 Freising, Germany.|Bavarian Biomolecular Mass Spectrometry Center (BayBioMS), Technical University of Munich , 85354 Freising, Germany.
authors: Bernhard
authors: B
authors: Kuster
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry / Q000378:metabolism; D045744:Cell Line, Tumor; D002986:Clinical Trials as Topic; D055808:Drug Discovery / Q000379:methods*; D006801:Humans; D008024:Ligands; D008958:Models, Molecular; D011485:Protein Binding; D047428:Protein Kinase Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D040901:Proteomics / Q000379:methods*; D036104:Receptor, EphA2 / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism*
chemical_list: D000596:Amino Acids; D008024:Ligands; D047428:Protein Kinase Inhibitors; D036104:Receptor, EphA2