title: Activation of halophilic nucleoside diphosphate kinase by a non-ionic osmolyte, trimethylamine N-oxide.
abstract: The folding and activity of halophilic enzymes are believed to require the presence of salts at high concentrations. When the inactivated nucleoside diphosphate kinase (NDK) from extremely halophilic archaea was incubated with low salt media, no activity was regained over the course of 8 days. When it was incubated with approximately 2 M NaCl or 3 M KCl, however, it gradually regained activity. To our surprise, trimethylamine N-oxide (TMAO) also was able to induce activation at 4.0 M. The enzyme activity and secondary structure of refolded NDK in 4 M TMAO were comparable with those of the native NDK or the refolded NDK in 3.8 M NaCl. TMAO is not an electrolyte, meaning that the presence of concentrated salts is not an absolute requirement, and that charge shielding or ion binding is not a sole factor for the folding and activation of NDK. Although both NaCl and TMAO are effective in refolding NDK, the mechanism of their actions appears to be different: the effect of protein concentration and pH on refolding is qualitatively different between these two, and at pH 8.0 NDK could be refolded in the presence of 4 M TMAO only when low concentrations of NaCl are included.
mesh_terms: D002942:Circular Dichroism; D004789:Enzyme Activation / Q000187:drug effects; D019617:Halobacterium salinarum / Q000201:enzymology; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D008744:Methylamines / Q000494:pharmacology*; D009701:Nucleoside-Diphosphate Kinase / Q000737:chemistry* / Q000378:metabolism*; D011189:Potassium Chloride / Q000494:pharmacology; D011487:Protein Conformation / Q000187:drug effects; D011489:Protein Denaturation; D017510:Protein Folding; D012965:Sodium Chloride / Q000494:pharmacology; D013997:Time Factors; D014508:Urea / Q000494:pharmacology
chemical_list: D008744:Methylamines; D012965:Sodium Chloride; D011189:Potassium Chloride; D014508:Urea; D009701:Nucleoside-Diphosphate Kinase; C005855:trimethyloxamine