title: Functional Role of Histidine in the Conserved His-x-Asp Motif in the Catalytic Core of Protein Kinases.
abstract: The His-x-Asp (HxD) motif is one of the most conserved structural components of the catalytic core of protein kinases; however, the functional role of the conserved histidine is unclear. Here we report that replacement of the HxD-histidine with Arginine or Phenylalanine in Aurora A abolishes both the catalytic activity and auto-phosphorylation, whereas the Histidine-to-tyrosine impairs the catalytic activity without affecting its auto-phosphorylation. Comparisons of the crystal structures of wild-type (WT) and mutant Aurora A demonstrate that the impairment of the kinase activity is accounted for by (1) disruption of the regulatory spine in the His-to-Arg mutant, and (2) change in the geometry of backbones of the Asp-Phe-Gly (DFG) motif and the DFG-1 residue in the His-to-Tyr mutant. In addition, bioinformatics analyses show that the HxD-histidine is a mutational hotspot in tumor tissues. Moreover, the H174R mutation of the HxD-histidine, in the tumor suppressor LKB1 abrogates the inhibition of anchorage-independent growth of A549 cells by WT LKB1. Based on these data, we propose that the HxD-histidine is involved in a conserved inflexible organization of the catalytic core that is required for the kinase activity. Mutation of the HxD-histidine may also be involved in the pathogenesis of some diseases including cancer.
authors.affiliation: National Center for Drug Screening and State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 189 Guo Shoujing Road, Shanghai 201203, China.
authors.forename: Lun
authors.initials: L
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 320 Yueyang Road, Shanghai 200031, China.
authors.forename: Jian-Chuan
authors.initials: JC
authors.lastname: Wang
authors.affiliation: National Center for Drug Screening and State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 189 Guo Shoujing Road, Shanghai 201203, China.
authors.forename: Li
authors.initials: L
authors.lastname: Hou
authors.affiliation: National Center for Drug Screening and State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 189 Guo Shoujing Road, Shanghai 201203, China.
authors.forename: Peng-Rong
authors.initials: PR
authors.lastname: Cao
authors.affiliation: School of Life Sciences, East China Normal University, 3663 North Zhongshan Road, Shanghai 200062, China.
authors.forename: Li
authors.initials: L
authors.lastname: Wu
authors.affiliation: Drug Discovery and Design Center, State Key Laboratory of Drug Research and Chinese Academy of Sciences Key Laboratory of Receptor Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 555 Zu Chongzhi Road, Shanghai 201203, China.
authors.forename: Qian-Sen
authors.initials: QS
authors.lastname: Zhang
authors.affiliation: Drug Discovery and Design Center, State Key Laboratory of Drug Research and Chinese Academy of Sciences Key Laboratory of Receptor Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 555 Zu Chongzhi Road, Shanghai 201203, China.
authors.forename: Huai-Yu
authors.initials: HY
authors.lastname: Yang
authors.affiliation: National Center for Drug Screening and State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 189 Guo Shoujing Road, Shanghai 201203, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Zang
authors.affiliation: State Key Laboratory of Molecular Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 320 Yueyang Road, Shanghai 200031, China.
authors.forename: Jian-Ping
authors.initials: JP
authors.lastname: Ding
authors.affiliation: National Center for Drug Screening and State Key Laboratory of Drug Research, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, 189 Guo Shoujing Road, Shanghai 201203, China.|School of Life Sciences, East China Normal University, 3663 North Zhongshan Road, Shanghai 200062, China.
authors.forename: Jia
authors.initials: J
authors.lastname: Li
mesh_terms: D000091162:AMP-Activated Protein Kinase Kinases; D020816:Amino Acid Motifs; D001120:Arginine / Q000235:genetics; D064096:Aurora Kinase A / Q000737:chemistry / Q000235:genetics; D020134:Catalytic Domain; D045744:Cell Line, Tumor; D017124:Conserved Sequence; D006639:Histidine / Q000378:metabolism*; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D008958:Models, Molecular; D009154:Mutation / Q000235:genetics; D011487:Protein Conformation; D011494:Protein Kinases / Q000737:chemistry* / Q000378:metabolism*; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry / Q000235:genetics; D013329:Structure-Activity Relationship; D014443:Tyrosine / Q000235:genetics; D014961:X-Ray Diffraction
chemical_list: D014443:Tyrosine; D006639:Histidine; D001120:Arginine; D011494:Protein Kinases; D064096:Aurora Kinase A; D017346:Protein Serine-Threonine Kinases; C109790:STK11 protein, human; D000091162:AMP-Activated Protein Kinase Kinases