title: Discovery of a novel allosteric inhibitor-binding site in ERK5: comparison with the canonical kinase hinge ATP-binding site.
abstract: MAP kinases act as an integration point for multiple biochemical signals and are involved in a wide variety of cellular processes such as proliferation, differentiation, regulation of transcription and development. As a member of the MAP kinase family, ERK5 (MAPK7) is involved in the downstream signalling pathways of various cell-surface receptors, including receptor tyrosine kinases and G protein-coupled receptors. In the current study, five structures of the ERK5 kinase domain co-crystallized with ERK5 inhibitors are reported. Interestingly, three of the compounds bind at a novel allosteric binding site in ERK5, while the other two bind at the typical ATP-binding site. Binding of inhibitors at the allosteric site is accompanied by displacement of the P-loop into the ATP-binding site and is shown to be ATP-competitive in an enzymatic assay of ERK5 kinase activity. Kinase selectivity data show that the most potent allosteric inhibitor exhibits superior kinase selectivity compared with the two inhibitors that bind at the canonical ATP-binding site. An analysis of these structures and comparison with both a previously published ERK5-inhibitor complex structure (PDB entry 4b99) and the structures of three other kinases (CDK2, ITK and MEK) in complex with allosteric inhibitors are presented.
authors.affiliation: Chemistry Innovation Centre, Discovery Sciences, AstraZeneca R&D Mölndal, 431 83 Mölndal, Sweden.
authors.forename: Hongming
authors.initials: H
authors.lastname: Chen
authors.affiliation: Structure and Biophysics, Discovery Sciences, AstraZeneca R&D Alderley Park, Macclesfield SK10 4TG, England.
authors.forename: Julie
authors.initials: J
authors.lastname: Tucker
authors.affiliation: Innovation Centre China, AstraZeneca Asia and Emerging Markets iMed, Shanghai 201203, People's Republic of China.
authors.forename: Xiaotao
authors.initials: X
authors.lastname: Wang
authors.affiliation: Innovation Centre China, AstraZeneca Asia and Emerging Markets iMed, Shanghai 201203, People's Republic of China.
authors.forename: Paul R
authors.initials: PR
authors.lastname: Gavine
authors.affiliation: Structure and Biophysics, Discovery Sciences, AstraZeneca R&D Alderley Park, Macclesfield SK10 4TG, England.
authors.forename: Chris
authors.initials: C
authors.lastname: Phillips
authors.affiliation: Proteros biostructures GmbH, Bunsenstrasse 7a, 82152 Martinsried, Germany.
authors.forename: Martin A
authors.initials: MA
authors.lastname: Augustin
authors.affiliation: Proteros biostructures GmbH, Bunsenstrasse 7a, 82152 Martinsried, Germany.
authors.forename: Patrick
authors.initials: P
authors.lastname: Schreiner
authors.affiliation: Proteros biostructures GmbH, Bunsenstrasse 7a, 82152 Martinsried, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Steinbacher
authors.affiliation: Screening Sciences, Discovery Sciences, AstraZeneca R&D Alderley Park, Macclesfield SK10 4TG, England.
authors.forename: Marian
authors.initials: M
authors.lastname: Preston
authors.affiliation: Structure and Biophysics, Discovery Sciences, AstraZeneca R&D Alderley Park, Macclesfield SK10 4TG, England.
authors.forename: Derek
authors.initials: D
authors.lastname: Ogg
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000494:Allosteric Regulation / Q000187:drug effects; D000495:Allosteric Site / Q000187:drug effects; D001665:Binding Sites / Q000187:drug effects; D018360:Crystallography, X-Ray; D006801:Humans; D048054:Mitogen-Activated Protein Kinase 7 / Q000037:antagonists & inhibitors* / Q000737:chemistry* / Q000378:metabolism; D062105:Molecular Docking Simulation; D047428:Protein Kinase Inhibitors / Q000737:chemistry / Q000494:pharmacology*
chemical_list: D047428:Protein Kinase Inhibitors; D000255:Adenosine Triphosphate; D048054:Mitogen-Activated Protein Kinase 7
keywords: ERK5 kinase; MAPK7 kinase; allosteric kinase inhibitor; crystal structure