Evidence for major structural changes in subunit C of the vacuolar ATPase due to nucleotide binding

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• 作者：Armbr脙脙ster ; Andrea ; Hohn ; Christina ; Hermesdorf ; Anne ; Schumacher ; Karin ; B脙脙rsch ; Michael ; et. al.
• 关键词：Vacuolar-type ATPase ; V₁V₀ ATPase ; V₁ ATPase ; Vma5p ; VHA-C ; Photoaffinity labeling ; Fluorescence correlation spectroscopy
• 刊名：FEBS Letters
• 出版年：2005
• 期刊代码：70_00145793
• 类别：bio
• 出版时间：March 28, 2005
• 卷：579
• 期：9
• 页码：1961-1967
• 文件大小：314 K

摘要

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.