@article{4f1d4b84de0b437ab67a1d1d50b59769,
title = "Functional analysis of conserved cysteine residues in the catalytic subunit of the yeast vacuolar H+-ATPase",
abstract = "The A subunit of the yeast vacuolar ATPase contains three highly conserved cysteines: Cys-261, Cys-284, and Cys-538. Cys-261 is located within the nucleotide-binding P-loop. Each of the conserved cysteines, and one nonconserved cysteine, Cys-254, were altered to serine by site-directed mutagenesis, and the effects on growth at pH 7.5 were determined. The Cys-254 → Ser, Cys-261 → Ser and the double mutants all grew at pH 7.5 and contained nitrate- and bafilomycin-sensitive ATPase activity. However, the ATPase activities of the Cys-261 → Ser and the double mutants were insensitive to the sulfhydryl group inhibitor, N-ethylmaleimide, demonstrating that Cys-261 is the site of inhibition by N-ethylmaleimide. Changing either Cys-284 or Cys-538 to serine prevented growth at pH 7.5. Cys-284 and Cys-538 thus appear to be essential cysteine residues which are required either for assembly or catalysis.",
keywords = "ATPase, Cysteine, H-, N-Ethylmaleimide, Proton pump, Vacuole, vacuolar",
author = "Lincoln Taiz and Hannah Nelson and Keith Maggert and Louis Morgan and Brad Yatabe and Taiz, {Saundra Lee} and Bernard Rubinstein and Nathan Nelson",
note = "Funding Information: This research was supported in part by Grant DE-FGO3-84ER13245 from the U.S. Department of Energy to L.T. The authors are indebted to Dr. Tom Stevens of the University of Oregon for generously providing the original plasmid containing the spacer-deleted A subunit gene which was utilized in preliminary experiments. We also wish to thank Drs. B.J. Bowman, E.J. Bowman, and J.P. Gogarten for helpful discussions.",
year = "1994",
month = sep,
day = "14",
doi = "10.1016/0005-2736(94)90315-8",
language = "אנגלית",
volume = "1194",
pages = "329--334",
journal = "BBA - Biomembranes",
issn = "0005-2736",
publisher = "Elsevier B.V.",
number = "2",
}