Mitochondrial F0F1 H+‐ATP synthase Characterization of F0 components involved in H+ translocation

FEBS Letters

Capozza, Giuseppe¹  Jirillo, Emilio¹  Zanotti, Franco¹  Colaianni, Gina¹  Guerrieri, Ferruccio¹  Houštěk, Josef¹  Papa, Sergio¹ 

[1] Institute of Medical Biochemistry and Chemistry, Centre for the Study of Mitochondria and Energy Metabolism, CNR and Institute of Immunology, University of Bari, Bari, Italy

关键词: Dicyclohexylcarbodiimide;    F0;    F1;    H+ transporting ATP synthase;    Oligomycin;    Proton translocation;    CHAPS;    3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate;    ESMP;    submitochondrial particles prepared in the presence of EDTA;    USMP;    submitochondrial particles devoid of F1 (see section 2);    SDS-PAGE;    SDS-polyacrylamide gel electrophoresis;    Enzyme: F0F1;    ATP synthase (EC 3.6.1.34);   

DOI  :  10.1016/0014-5793(89)80685-4

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The membrane F₀, sector of mitochondrial ATP synthase complex was rapidly isolated by direct extraction with CHAPS from F₁-depleted submitochondrial particles. The preparation thus obtained is stable and can be reconstituted in artificial phospholipid membranes to result in oligomycin-sensitive proton conduction, or recombined with purified F₁ to give the oligomycin-sensitive F₀F₁-ATPase complex. The F₀ preparation and constituent polypeptides were characterized by SDS-polyacrylamide gel electrophoresis and immunoblot analysis. The functional role of F₀ polypeptides was examined by means of trypsin digestion and reconstitution studies. It is shown that, in addition to the 8 kDa DCCD-binding protein, the nuclear encoded protein [(1987) J. Mol. Biol. 197, 89–100], characterized as an intrinsic component of F₀, (F₀I, PVP protein [(1967) J. Biol. Chem. 242, 2547–2551]) is involved in H⁺ translocation and the sensitivity of this process to the F₀ inhibitors, DCCD and oligomycin.

【 授权许可】

Unknown