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Identification of Subunits a, b, andc 1 from Acetobacterium woodiiNa+-F1F0-ATPase

2000; Elsevier BV; Volume: 275; Issue: 43 Linguagem: Inglês

10.1074/jbc.m005134200

1083-351X

Sascha Aufurth, Hermann Schägger, Volker Müller,

Ion Transport and Channel Regulation

The Na + -F 1 F 0 -ATPase operon of Acetobacterium woodii was recently shown to contain, among eleven atp genes, those genes that encode subunit a and b , a gene encoding a 16-kDa proteolipid (subunit c 1 ), and two genes encoding 8-kDa proteolipids (subunits c 2 and c 3 ). Because subunits a , b , and c 1 were not found in previous enzyme preparations, we re-determined the subunit composition of the enzyme. The genes were overproduced, and specific antibodies were raised. Western blots revealed that subunits a , b, and c 1 are produced and localized in the cytoplasmic membrane. Membrane protein complexes were solubilized by dodecylmaltoside and separated by blue native-polyacrylamide gel electrophoresis, and the ATPase subunits were resolved by SDS-polyacrylamide gel electrophoresis. N-terminal sequence analyses revealed the presence of subunits a , c 2 , c 3 , b , δ, α, γ, β, and ε. Biochemical and immunological analyses revealed that subunits c 1 , c 2 , and c 3 are all part of the c -oligomer, the first of a F 1 F 0 -ATPase that contains 8- and 16-kDa proteolipids.