Bacterial [Na.sup.+]-translocating ferredoxin: [NAD.sup.+] oxidoreductase

The anaerobic acetogenic bacterium Acetobacterium woodii carries out a unique type of [Na.sup.+]-motive, anaerobic respiration with caffeate as electron acceptor, termed 'caffeate respiration.' Central, and so far the only identified membrane-bound reaction in this respiration pathway, is a ferredoxin:[NAD.sup.+] oxidoreductase (Fno) activity. Here we show that inverted membrane vesicles of A. woodii couple electron transfer from reduced ferredoxin to [NAD.sup.+] with the transport of [Na.sup.+] from the outside into the lumen of the vesicles. [Na.sup.+] transport was electrogenic, and accumulation was inhibited by sodium ionophores but not protonophores, demonstrating a direct coupling of Fno activity to [Na.sup.+] transport. Results from inhibitor studies are consistent with the hypothesis that Fno activity coupled to [Na.sup.+] translocation is catalyzed by the Rnf complex, a membranebound, iron--sulfur and flavin-containing electron transport complex encoded by many bacterial and some archaeal genomes. Fno is a unique type of primary [Na.sup.+] pump and represents an early evolutionary mechanism of energy conservation that expands the redox range known to support life. In addition, it explains the lifestyle of many anaerobic bacteria and gives a mechanistic explanation for the enigma of the energetic driving force for the endergonic reduction of ferredoxin with NADH plus [H.sup.+] as reductant in a number of aerobic bacteria. Acetobacterium | anaerobic respiration | electron transport | [Na.sup.+] pump | Rnf doi/ 10.1073/pnas.1010318107