The ability of subunit C of eukaryotic V-ATPases to bind ADP and ATP is demonstrated by photoaffinity labeling and fluorescence correlation spectroscopy (FCS). Quantitation of the photoaffinity and the FCS data indicate that the ATP-analogues bind more weakly to subunit C than the ADP-analogues. Site-directed mutagenesis and N-terminal sequencing of subunit C from Arabidopsis (VHA-C) and yeast (Vma5p) have been used to map the C-terminal region of subunit C as the nucleotide-binding site. Tryptophan fluorescence quenching and decreased susceptibility to tryptic digestion of subunit C after binding of different nucleotides provides evidence for structural changes in this subunit caused by nucleotide-binding. © 2005 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.
Armbrüster, A., Hohn, C., Hermesdorf, A., Schumacher, K., Börsch, M., & Grüber, G. (2005). Evidence for major structural changes in subunit C of the vacuolar ATPase due to nucleotide binding. FEBS Letters, 579(9), 1961–1967. https://doi.org/10.1016/j.febslet.2005.02.042