title: Kinase regulation by hydrophobic spine assembly in cancer.
abstract: A new model of kinase regulation based on the assembly of hydrophobic spines has been proposed. Changes in their positions can explain the mechanism of kinase activation. Here, we examined mutations in human cancer for clues about the regulation of the hydrophobic spines by focusing initially on mutations to Phe. We identified a selected number of Phe mutations in a small group of kinases that included BRAF, ABL1, and the epidermal growth factor receptor. Testing some of these mutations in BRAF, we found that one of the mutations impaired ATP binding and catalytic activity but promoted noncatalytic allosteric functions. Other Phe mutations functioned to promote constitutive catalytic activity. One of these mutations revealed a previously underappreciated hydrophobic surface that functions to position the dynamic regulatory αC-helix. This supports the key role of the C-helix as a signal integration motif for coordinating multiple elements of the kinase to create an active conformation. The importance of the hydrophobic space around the αC-helix was further tested by studying a V600F mutant, which was constitutively active in the absence of the negative charge that is associated with the common V600E mutation. Many hydrophobic mutations strategically localized along the C-helix can thus drive kinase activation.
authors.affiliation: Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, Missouri, USA Howard Hughes Medical Institute, Washington University School of Medicine, St. Louis, Missouri, USA.
authors.forename: Jiancheng
authors.identifier: http://orcid.org/0000-0003-1615-0030
authors.initials: J
authors.lastname: Hu
authors.affiliation: Department of Pharmacology, University of California, San Diego, La Jolla, California, USA.
authors.forename: Lalima G
authors.initials: LG
authors.lastname: Ahuja
authors.affiliation: Department of Pharmacology, University of California, San Diego, La Jolla, California, USA Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, California, USA.
authors.forename: Hiruy S
authors.initials: HS
authors.lastname: Meharena
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Georgia, Athens, Georgia, USA.
authors.forename: Natarajan
authors.initials: N
authors.lastname: Kannan
authors.affiliation: Department of Pharmacology, University of California, San Diego, La Jolla, California, USA Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, California, USA Howard Hughes Medical Institute, University of California, San Diego, La Jolla, California, USA.
authors.forename: Alexandr P
authors.initials: AP
authors.lastname: Kornev
authors.affiliation: Department of Pharmacology, University of California, San Diego, La Jolla, California, USA Department of Chemistry and Biochemistry, University of California, San Diego, La Jolla, California, USA Howard Hughes Medical Institute, University of California, San Diego, La Jolla, California, USA staylor@ucsd.edu shaw@pathbox.wustl.edu.
authors.forename: Susan S
authors.initials: SS
authors.lastname: Taylor
authors.affiliation: Department of Pathology and Immunology, Washington University School of Medicine, St. Louis, Missouri, USA Howard Hughes Medical Institute, Washington University School of Medicine, St. Louis, Missouri, USA staylor@ucsd.edu shaw@pathbox.wustl.edu.
authors.forename: Andrey S
authors.initials: AS
authors.lastname: Shaw
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000495:Allosteric Site; D002384:Catalysis; D066246:ErbB Receptors / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic; D057809:HEK293 Cells; D006639:Histidine / Q000737:chemistry; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D008715:Methionine / Q000737:chemistry; D008958:Models, Molecular; D009154:Mutation; D009369:Neoplasms / Q000201:enzymology*; D010770:Phosphotransferases / Q000502:physiology*; D017433:Protein Structure, Secondary; D048493:Proto-Oncogene Proteins B-raf / Q000235:genetics; D016315:Proto-Oncogene Proteins c-abl / Q000235:genetics
chemical_list: D006639:Histidine; D000255:Adenosine Triphosphate; D008715:Methionine; D010770:Phosphotransferases; D066246:ErbB Receptors; D016315:Proto-Oncogene Proteins c-abl; C482119:BRAF protein, human; D048493:Proto-Oncogene Proteins B-raf