Effects of mutations at Gly114 on the stability and refolding of Haloarchaeal nucleoside diphosphate kinase in low salt solution.
We have shown before that mutation of Gly114 to Arg enhances folding of hexameric nucleoside diphosphate kinase (HsNDK) from Halobacterium salinarum. In this study, we constructed three mutant forms, Gly114Lys (G114K), Gly114Ser (G114S) and Gly114Asp (G114D), to further clarify the role residue 114 plays in the stability and folding of HsNDK. While expression of G114D mutant resulted in inactive enzyme, other mutant HsNDKs were successfully expressed in active form. The G114K mutant, similar to Gly114Arg (G114R) mutant, refolded in 1M NaCl after heat-denaturation, under which the wild-type HsNDK and G114S proteins showed no refolding.
Ishibashi
Matsujiro
M
Faculty of Agriculture, Kagoshima University, 1-21-24 Korimoto, Kagoshima 890-0065, Japan. matu@ms.kagoshima-u.ac.jp
Iwasa
Tatsuya
T
Kumeda
Kouko
K
Arakawa
Tsutomu
T
Tokunaga
Masao
M
D004795:Enzyme Stability / Q000187:drug effects; D005998:Glycine / Q000235:genetics*; D019617:Halobacterium salinarum / Q000201:enzymology*; D009154:Mutation; D009701:Nucleoside-Diphosphate Kinase / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D017510:Protein Folding / Q000187:drug effects; D020673:Protein Renaturation / Q000187:drug effects*; D020836:Protein Structure, Quaternary / Q000187:drug effects; D012965:Sodium Chloride / Q000494:pharmacology*; D012996:Solutions; D013696:Temperature
D012996:Solutions; D012965:Sodium Chloride; D009701:Nucleoside-Diphosphate Kinase; D005998:Glycine