Spannagel C, Vaillier J, Arselin G, Graves P V, Velours J
Institut de Biochimie et Génétique Cellulaires du CNRS, Université Victor Segalen, Bordeaux 2, France.
Eur J Biochem. 1997 Aug 1;247(3):1111-7. doi: 10.1111/j.1432-1033.1997.01111.x.
The subunit f of the yeast F1F0ATP synthase has been isolated from the purified enzyme. Amino acid composition, protein and peptide sequencing were performed. The data are in agreement with the sequence of the predicted product of the gene D9481.21 identified on the Saccharomyces cerevisiae chromosome IV. A 303-bp open reading frame encoding a 101-amino acid polypeptide is described. The deduced amino acid sequence from the ATP17 gene is 6 amino acids longer than the mature protein, which displays a molecular mass of 10567 Da. The protein is basic with a short hydrophobic segment located in the C-terminal part of the subunit. Subunit f remained associated with other F0 subunits upon sodium bromide treatment of the whole enzyme. A null mutant was constructed. The disrupted strain was unable to grow on glycerol medium and the mutation was recessive; rho- cells arose spontaneously. The null mutant mitochondria were devoid of oligomycin-sensitive ATPase, but still contained an active F1, while the subunits f, 6 and 8 were absent.
酵母F1F0ATP合酶的亚基f已从纯化的酶中分离出来。进行了氨基酸组成分析、蛋白质和肽测序。数据与在酿酒酵母IV号染色体上鉴定出的基因D9481.21的预测产物序列一致。描述了一个编码101个氨基酸多肽的303 bp开放阅读框。从ATP17基因推导的氨基酸序列比成熟蛋白长6个氨基酸，成熟蛋白的分子量为10567 Da。该蛋白呈碱性，在亚基的C末端部分有一个短的疏水片段。用溴化钠处理全酶后，亚基f仍与其他F0亚基结合。构建了一个缺失突变体。缺失突变株在甘油培养基上无法生长，且该突变是隐性的；rho-细胞自发产生。缺失突变体的线粒体缺乏对寡霉素敏感的ATP酶，但仍含有活性F1，而亚基f、6和8缺失。
