title: Structure of the Human Protein Kinase ZAK in Complex with Vemurafenib.
abstract: The mixed lineage kinase ZAK is a key regulator of the MAPK pathway mediating cell survival and inflammatory response. ZAK is targeted by several clinically approved kinase inhibitors, and inhibition of ZAK has been reported to protect from doxorubicin-induced cardiomyopathy. On the other hand, unintended targeting of ZAK has been linked to severe adverse effects such as the development of cutaneous squamous cell carcinoma. Therefore, both specific inhibitors of ZAK, as well as anticancer drugs lacking off-target activity against ZAK, may provide therapeutic benefit. Here, we report the first crystal structure of ZAK in complex with the B-RAF inhibitor vemurafenib. The cocrystal structure displayed a number of ZAK-specific features including a highly distorted P loop conformation enabling rational inhibitor design. Positional scanning peptide library analysis revealed a unique substrate specificity of the ZAK kinase including unprecedented preferences for histidine residues at positions -1 and +2 relative to the phosphoacceptor site. In addition, we screened a library of clinical kinase inhibitors identifying several inhibitors that potently inhibit ZAK, demonstrating that this kinase is commonly mistargeted by currently used anticancer drugs.
authors.affiliation: Structural Genomics Consortium (SGC), Nuffield Department of Medicine, University of Oxford , Oxford, OX37DQ, United Kingdom.|Target Discovery Institute (TDI), Nuffield Department of Medicine, University of Oxford , Oxford, OX37FZ, United Kingdom.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Mathea
authors.affiliation: Structural Genomics Consortium (SGC), Nuffield Department of Medicine, University of Oxford , Oxford, OX37DQ, United Kingdom.
authors.forename: Kamal R
authors.initials: KR
authors.lastname: Abdul Azeez
authors.affiliation: Structural Genomics Consortium (SGC), Nuffield Department of Medicine, University of Oxford , Oxford, OX37DQ, United Kingdom.|Target Discovery Institute (TDI), Nuffield Department of Medicine, University of Oxford , Oxford, OX37FZ, United Kingdom.
authors.forename: Eidarus
authors.initials: E
authors.lastname: Salah
authors.affiliation: Structural Genomics Consortium (SGC), Nuffield Department of Medicine, University of Oxford , Oxford, OX37DQ, United Kingdom.|Target Discovery Institute (TDI), Nuffield Department of Medicine, University of Oxford , Oxford, OX37FZ, United Kingdom.
authors.forename: Cynthia
authors.initials: C
authors.lastname: Tallant
authors.affiliation: Target Discovery Institute (TDI), Nuffield Department of Medicine, University of Oxford , Oxford, OX37FZ, United Kingdom.
authors.forename: Finn
authors.initials: F
authors.lastname: Wolfreys
authors.affiliation: Target Discovery Institute (TDI), Nuffield Department of Medicine, University of Oxford , Oxford, OX37FZ, United Kingdom.
authors.forename: Rebecca
authors.initials: R
authors.lastname: Konietzny
authors.affiliation: Target Discovery Institute (TDI), Nuffield Department of Medicine, University of Oxford , Oxford, OX37FZ, United Kingdom.
authors.forename: Roman
authors.initials: R
authors.lastname: Fischer
authors.affiliation: Department of Pharmacology, Yale University School of Medicine , New Haven, Connecticut 06520, United States.
authors.forename: Hua Jane
authors.initials: HJ
authors.lastname: Lou
authors.affiliation: Target Discovery Institute (TDI), Nuffield Department of Medicine, University of Oxford , Oxford, OX37FZ, United Kingdom.
authors.forename: Paul E
authors.initials: PE
authors.lastname: Brennan
authors.affiliation: Lead Discovery and Optimisation Support, Boehringer Ingelheim Pharma GmbH & Co KG , Biberach, 88400, Germany.
authors.forename: Gisela
authors.initials: G
authors.lastname: Schnapp
authors.affiliation: Lead Discovery and Optimisation Support, Boehringer Ingelheim Pharma GmbH & Co KG , Biberach, 88400, Germany.
authors.forename: Alexander
authors.initials: A
authors.lastname: Pautsch
authors.affiliation: Target Discovery Institute (TDI), Nuffield Department of Medicine, University of Oxford , Oxford, OX37FZ, United Kingdom.
authors.forename: Benedikt M
authors.initials: BM
authors.lastname: Kessler
authors.affiliation: Department of Pharmacology, Yale University School of Medicine , New Haven, Connecticut 06520, United States.
authors.forename: Benjamin E
authors.initials: BE
authors.lastname: Turk
authors.affiliation: Target Discovery Institute (TDI), Nuffield Department of Medicine, University of Oxford , Oxford, OX37FZ, United Kingdom.|Institute for Pharmaceutical Chemistry and Buchmann Institute for Molecular Life Sciences (BMLS), Johann Wolfgang Goethe University , Frankfurt am Main, 60438, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Knapp
mesh_terms: D000255:Adenosine Triphosphate / Q000031:analogs & derivatives / Q000737:chemistry; D002151:Calorimetry; D018360:Crystallography, X-Ray; D015195:Drug Design; D006801:Humans; D007211:Indoles / Q000737:chemistry*; D016350:Leucine Zippers; D020930:MAP Kinase Kinase Kinases; D015394:Molecular Structure; D009842:Oligopeptides / Q000138:chemical synthesis / Q000737:chemistry; D010750:Phosphoproteins / Q000737:chemistry; D010766:Phosphorylation; D047428:Protein Kinase Inhibitors / Q000737:chemistry*; D011494:Protein Kinases / Q000737:chemistry*; D048493:Proto-Oncogene Proteins B-raf / Q000037:antagonists & inhibitors / Q000737:chemistry; D013449:Sulfonamides / Q000737:chemistry*; D000077484:Vemurafenib
chemical_list: D007211:Indoles; D009842:Oligopeptides; D010750:Phosphoproteins; D047428:Protein Kinase Inhibitors; D013449:Sulfonamides; D000077484:Vemurafenib; C022571:adenosine 5'-O-(3-thiotriphosphate); D000255:Adenosine Triphosphate; D011494:Protein Kinases; D048493:Proto-Oncogene Proteins B-raf; D020930:MAP Kinase Kinase Kinases; C413047:MAP3K20 protein, human