Foster D L, Mosher M E, Futai M, Fillingame R H
J Biol Chem. 1980 Dec 25;255(24):12037-41.
The proton-translocating ATPase complex (F1F0) of Escherichia coli was purified after inductin of a lambda-transducing phage (lambda asn5) carrying the ATPase genes of th unc operon. ATPase activity of membranes prepared from the induced lambda-unc lysogen was 6-fold greater than the activity of membranes prepared from strains lacking the unc-transducing phage, confirming the report of Kanazawa et al. (1979) Proc. Natl. Acad. Sci. U. S. A. 76, 1126-1130). The F1F0-ATPase complex was purified in comparable yield from either enriched membranes or control membranes using a modification of the procedure reported by Foster and Fillingame ((1979) J. Biol. Chem. 254, 8230-8236). EAch of the eight subunits that had been reported as components of the F1F0 complex from wild type E. coli was overproduced in the lambda-unc lysogen. All eight subunits co-purified in the same stoichiometric proportion as in the complex purified from wild type E. coli. We conclude that all eight subunits are likely coded by the small segment of chromosomal DNA carried by the lambda-transducing phage. These experiments provide the first evidence that all eight polypeptides are authentic subunits of the ATPase complex rather than contaminants that fortuitously co-purify.
携带unc操纵子ATP酶基因的λ转导噬菌体（λasn5）诱导后，纯化了大肠杆菌的质子转运ATP酶复合物（F1F0）。由诱导的λ-unc溶原菌制备的膜的ATP酶活性比缺乏unc转导噬菌体的菌株制备的膜的活性高6倍，这证实了金泽等人（1979年，《美国国家科学院院刊》76卷，1126 - 1130页）的报道。使用福斯特和菲林盖姆（1979年，《生物化学杂志》254卷，8230 - 8236页）报道的方法的改进方法，从富集膜或对照膜中以相当的产量纯化了F1F0 - ATP酶复合物。据报道，野生型大肠杆菌F1F0复合物的八个亚基中的每一个在λ-unc溶原菌中都过量产生。所有八个亚基以与从野生型大肠杆菌纯化的复合物相同的化学计量比例共同纯化。我们得出结论，所有八个亚基可能由λ转导噬菌体携带的一小段染色体DNA编码。这些实验提供了第一个证据，即所有八个多肽都是ATP酶复合物的真实亚基，而不是偶然共同纯化的污染物。
