title: The interaction between residues 62 and 193 play a key role in activity and structural stability of arginine kinase.
abstract: The purpose of this study is to clarify that the amino acid residues (Asp62 and Arg193) are responsible for the activity and stability of arginine kinase (AK). The amino acid residues Asp62 (D62) and Arg193 (R193) are strictly conserved in monomeric AKs and form an ion pair in the transition state analogue complex. In this research, we replaced D62 with glutamate (E) or glycine (G) and R193 with lysine (K) or glycine (G). The mutants of D62E and R193K retained almost 90% of the wild-type activity, whereas D62G and R193G had a pronounced loss in activity. A detailed comparison was made between the physic-chemical properties and conformational changes of wild-type AK and the mutants by means of ultraviolet (UV) difference and fluorescence spectra. The results indicated that the conformation of all of the mutants had been changed and the stability in a urea solution was also reduced. We speculated that the hydrogen bond and electrostatic interactions formed between residues 62 and 193 play a key role in stabilizing the structure and mediating the synergism in substrate binding of arginine kinase from greasyback shrimp (Metapenaeus ensis).
authors.affiliation: Hubei Normal University, Huangshi, Hubei 435002, People's Republic of China.
authors.forename: Na
authors.initials: N
authors.lastname: Liu
authors.forename: Jin-Song
authors.initials: JS
authors.lastname: Wang
authors.forename: Wei-Dong
authors.initials: WD
authors.lastname: Wang
authors.forename: Ji-Cheng
authors.initials: JC
authors.lastname: Pan
mesh_terms: D000818:Animals; D001122:Arginine Kinase / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D004795:Enzyme Stability; D006860:Hydrogen Bonding; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D033561:Penaeidae / Q000201:enzymology; D011485:Protein Binding; D011487:Protein Conformation; D058767:Protein Unfolding
chemical_list: D001122:Arginine Kinase