The Na+-translocating NADH:quinone oxidoreductase (Na+-NQR) from Vibrio alginolyticus was inactivated by reactive oxygen species. Highest Na+-NQR activity was observed in anaerobically prepared membranes that exhibited 1 : 1 coupling of NADH oxidation and Q reduction activities ( UÆmg)1). Optical and EPR spectroscopy documented the presence of b-type cytochromes, a [2Fe)2S] cluster and an organic radical signal in anaerobically pre- pared membranes from V. alginolyticus.