Solubilization and reconstitution of the Na+-motive NADH oxidase activity from the marine bacterium Vibrio alginolyticus.
The Na+-motive NADH oxidase activity from Vibrio alginolyticus was extracted with octylglucoside and reconstituted into liposomes by dilution. On the addition of NADH, the reconstituted proteoliposomes generated delta psi (inside positive) and delta pH (inside alkaline) in the presence of a proton conductor CCCP, and accumulated Na+ in the presence of valinomycin. These results indicate that the NADH oxidase activity, reconstituted in opposite orientation, leads to the generation of an electrochemical potential of Na+ by the influx of Na+.
D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone / Q000494:pharmacology; D004563:Electrochemistry; D006863:Hydrogen-Ion Concentration; D007473:Ion Channels / Q000378:metabolism; D008081:Liposomes / Q000378:metabolism; D009097:Multienzyme Complexes / Q000302:isolation & purification / Q000378:metabolism*; D009243:NAD / Q000494:pharmacology; D009247:NADH, NADPH Oxidoreductases / Q000302:isolation & purification / Q000378:metabolism*; D012964:Sodium / Q000378:metabolism* / Q000494:pharmacology; D014451:Ubiquinone / Q000494:pharmacology; D014634:Valinomycin / Q000494:pharmacology; D014733:Vibrio / Q000201:enzymology*
D007473:Ion Channels; D008081:Liposomes; D009097:Multienzyme Complexes; D009243:NAD; D014451:Ubiquinone; D014634:Valinomycin; D002258:Carbonyl Cyanide m-Chlorophenyl Hydrazone; D012964:Sodium; C023107:NADH oxidase; D009247:NADH, NADPH Oxidoreductases; C025203:Ubiquinone Q1