title: Insights into the Phosphoryl Transfer Mechanism of Human Ubiquitous Mitochondrial Creatine Kinase.
abstract: Human ubiquitous mitochondrial creatine kinase (uMtCK) is responsible for the regulation of cellular energy metabolism. To investigate the phosphoryl-transfer mechanism catalyzed by human uMtCK, in this work, molecular dynamic simulations of uMtCK∙ATP-Mg2+∙creatine complex and quantum mechanism calculations were performed to make clear the puzzle. The theoretical studies hereof revealed that human uMtCK utilizes a two-step dissociative mechanism, in which the E227 residue of uMtCK acts as the catalytic base to accept the creatine guanidinium proton. This catalytic role of E227 was further confirmed by our assay on the phosphatase activity. Moreover, the roles of active site residues in phosphoryl transfer reaction were also identified by site directed mutagenesis. This study reveals the structural basis of biochemical activity of uMtCK and gets insights into its phosphoryl transfer mechanism.
authors.affiliation: Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences &Peking Union Medical College, Beijing 100050, China.
authors.forename: Quanjie
authors.initials: Q
authors.lastname: Li
authors.affiliation: Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences &Peking Union Medical College, Beijing 100050, China.
authors.forename: Shuai
authors.initials: S
authors.lastname: Fan
authors.affiliation: Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences &Peking Union Medical College, Beijing 100050, China.
authors.forename: Xiaoyu
authors.initials: X
authors.lastname: Li
authors.affiliation: Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences &Peking Union Medical College, Beijing 100050, China.
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Jin
authors.affiliation: Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences &Peking Union Medical College, Beijing 100050, China.
authors.forename: Weiqing
authors.initials: W
authors.lastname: He
authors.affiliation: Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences &Peking Union Medical College, Beijing 100050, China.
authors.forename: Jinming
authors.initials: J
authors.lastname: Zhou
authors.affiliation: Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences &Peking Union Medical College, Beijing 100050, China.
authors.forename: Shan
authors.initials: S
authors.lastname: Cen
authors.affiliation: Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences &Peking Union Medical College, Beijing 100050, China.
authors.forename: ZhaoYong
authors.initials: Z
authors.lastname: Yang
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D019943:Amino Acid Substitution; D001665:Binding Sites; D020134:Catalytic Domain; D003401:Creatine / Q000737:chemistry / Q000378:metabolism; D003402:Creatine Kinase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D052281:Creatine Kinase, Mitochondrial Form / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D019791:Guanidine / Q000737:chemistry; D006801:Humans; D008274:Magnesium / Q000378:metabolism; D056004:Molecular Dynamics Simulation; D016297:Mutagenesis, Site-Directed
chemical_list: D000255:Adenosine Triphosphate; C000627607:CKMT1B protein, human; D003402:Creatine Kinase; D052281:Creatine Kinase, Mitochondrial Form; D008274:Magnesium; D019791:Guanidine; D003401:Creatine