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A single Gly114Arg mutation stabilizes the hexameric subunit assembly and changes the substrate specificity of halo‐archaeal nucleoside diphosphate kinase
Author(s) -
Ishibashi Matsujiro,
Tatsuda Shuhei,
Izutsu Ken-ichi,
Kumeda Kouko,
Arakawa Tsutomu,
Tokunaga Masao
Publication year - 2007
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2007.07.042
Subject(s) - mutant , protein subunit , nucleoside diphosphate kinase , biochemistry , escherichia coli , chemistry , nucleotide , residue (chemistry) , enzyme , mutation , nucleoside , kinase , nucleoside triphosphate , biology , microbiology and biotechnology , gene
Nucleoside diphosphate kinase from extremely halophilic archaeon (HsNDK) requires above 2 M NaCl concentration for in vitro refolding. Here an attempt was made to isolate mutations that allow HsNDK to refold in low salt media. Such a screening resulted in isolation of an HsNDK mutant, Gly114Arg, which efficiently refolded in the presence of 1 M NaCl. This mutant, unlike the wild type enzyme, was expressed in Escherichia coli as an active form. The residue 114 is in close proximity to Glu155 of the neighboring subunit in the three dimensional hexameric structure of the HsNDK. It is thus possible that the attractive electrostatic interactions occur between Arg114 and Glu155 in the mutant HsNDK, stabilizing the hexameric subunit assembly.