title: Mutation of the conserved Asp122 in the linker impedes creatine kinase reactivation and refolding.
abstract: Creatine kinase (CK), a key enzyme in maintaining the intracellular energetic homeostasis, contains two domains connected by a long linker. In this research,we found that the mutations of the conserved Asp122 in the linker slightly affected CK activity, structure and stability. The hydrogen bonding and the ion pair contributed 2-5 kJ/mol to the conformational stability of CK. Interestingly, the ability of CK reactivation from the denatured state was completely removed by the mutations. These results suggested that the electrostatic interactions were crucial to the action of the linker in CK reactivation.
authors.affiliation: Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing, China.
authors.forename: Yan-Ming
authors.initials: YM
authors.lastname: Liu
authors.forename: Shan
authors.initials: S
authors.lastname: Feng
authors.forename: Xiao-Lan
authors.initials: XL
authors.lastname: Ding
authors.forename: Chi-Fei
authors.initials: CF
authors.lastname: Kang
authors.forename: Yong-Bin
authors.initials: YB
authors.lastname: Yan
mesh_terms: D019943:Amino Acid Substitution / Q000187:drug effects; D000818:Animals; D001224:Aspartic Acid / Q000235:genetics*; D017124:Conserved Sequence; D003402:Creatine Kinase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D004789:Enzyme Activation / Q000187:drug effects; D004795:Enzyme Stability / Q000187:drug effects; D019791:Guanidine / Q000494:pharmacology; D050505:Mutant Proteins / Q000737:chemistry / Q000378:metabolism; D009154:Mutation / Q000235:genetics*; D011489:Protein Denaturation / Q000187:drug effects; D017510:Protein Folding / Q000187:drug effects; D020836:Protein Structure, Quaternary; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011817:Rabbits; D013329:Structure-Activity Relationship; D013696:Temperature
chemical_list: D050505:Mutant Proteins; D001224:Aspartic Acid; D003402:Creatine Kinase; D019791:Guanidine