title: Interaction of hexa-His tag with acidic amino acids results in facilitated refolding of halophilic nucleoside diphosphate kinase.
abstract: We have previously reported that amino-terminal extension sequence containing hexa-His facilitated refolding and assembly of hexameric nucleoside diphosphate kinase from extremely halophilic archaeon Halobacterium salinarum (NDK). In this study, we made various mutations in both the tag sequence and within NDK molecule. SerNDK, in which hexa-His was replaced with hexa-Ser, showed no facilitated folding. In addition, HisD58GD63G, in which both Asp58 and Asp63 in NDK were replaced with Gly, also showed no refolding enhancement. These results suggest that hexa-His in His-tag interact cooperatively with either Asp58 or Asp63 or both. Furthermore, G114D mutant, which formed a dimer in low salt solution, was strongly stabilized by His-tag to form a stable hexamer.
authors.affiliation: Faculty of Agriculture, Kagoshima University, Kagoshima, Japan. matu@ms.kagoshima-u.ac.jp
authors.forename: Matsujiro
authors.initials: M
authors.lastname: Ishibashi
authors.forename: Keiko
authors.initials: K
authors.lastname: Ida
authors.forename: Shuhei
authors.initials: S
authors.lastname: Tatsuda
authors.forename: Tsutomu
authors.initials: T
authors.lastname: Arakawa
authors.forename: Masao
authors.initials: M
authors.lastname: Tokunaga
mesh_terms: D000595:Amino Acid Sequence; D024342:Amino Acids, Acidic / Q000378:metabolism*; D002942:Circular Dichroism; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability / Q000187:drug effects; D019617:Halobacterium salinarum / Q000187:drug effects / Q000201:enzymology*; D006639:Histidine / Q000737:chemistry / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration / Q000187:drug effects; D008958:Models, Molecular; D008969:Molecular Sequence Data; D050505:Mutant Proteins / Q000302:isolation & purification / Q000378:metabolism; D009701:Nucleoside-Diphosphate Kinase / Q000737:chemistry / Q000378:metabolism*; D009842:Oligopeptides / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding / Q000187:drug effects; D058849:Protein Refolding / Q000187:drug effects; D020836:Protein Structure, Quaternary; D012965:Sodium Chloride / Q000494:pharmacology; D013499:Surface Properties / Q000187:drug effects; D013696:Temperature
chemical_list: D024342:Amino Acids, Acidic; C471213:His-His-His-His-His-His; D050505:Mutant Proteins; D009842:Oligopeptides; D012965:Sodium Chloride; D006639:Histidine; D009701:Nucleoside-Diphosphate Kinase