Evidence for the presence of an F-type ATP synthase involved in sulfate respiration in Desulfovibrio vulgaris.
Using a library of genomic DNA from Desulfovibrio vulgaris Miyazaki F, a strict anaerobe, and two synthetic deoxyoligonucleotide probes designed for F-type ATPases, the genes for open reading frames (ORFs) 1 to 5 were cloned and sequenced. The predicted protein sequences of the gene products indicate that they are composed of 172, 488, 294, 471, and 134 amino acids, respectively, and that they share considerable identity at the amino acid level with delta, alpha, gamma, beta, and epsilon subunits found in other F-type ATPases, respectively. Furthermore, a component carrying ATPase activity was partially purified from the cytoplasmic membrane fraction of the D. vulgaris Miyazaki F cells. The N-terminal amino acid sequences of three major polypeptides separated by sodium dodecyl sulfate-12% polyacrylamide gel electrophoresis were identical to those of the products predicted by the sequences of ORF-2, ORF-3, and ORF-4, suggesting that an F-type ATPase is functioning in the D. vulgaris Miyazaki F cytoplasmic membrane. The amount of the F-type ATPase produced in the D. vulgaris Miyazaki F cells is similar to that in the Escherichia coli cells cultured aerobically. It indicates that the enzyme works as an ATP synthase in the D. vulgaris Miyazaki F cells in connection with sulfate respiration.
Ozawa
K
K
Department of Chemistry and Biotechnology, Faculty of Engineering, Yokohama National University, Hodogaya-ku, Yokohama 240-8501, Japan.
Meikari
T
T
Motohashi
K
K
Yoshida
M
M
Akutsu
H
H
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D016969:Desulfovibrio vulgaris / Q000201:enzymology / Q000235:genetics*; D004579:Electron Transport; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D006180:Proton-Translocating ATPases / Q000235:genetics* / Q000302:isolation & purification; D013431:Sulfates / Q000378:metabolism*
D013431:Sulfates; D006180:Proton-Translocating ATPases