Mitochondrial F0F1 H+-ATP synthase. Characterization of F0 components involved in H+ translocation.
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 [(1988) FEBS Lett. 237,9-14]) [corrected] is involved in H+ translocation and the sensitivity of this process to the F0 inhibitors, DCCD and oligomycin.
Guerrieri
F
F
Institute of Medical Biochemistry and Chemistry, Centre for the Study of Mitochondria and Energy Metabolism, CNR, Bari, Italy.
Capozza
G
G
Houstĕk
J
J
Zanotti
F
F
Colaianni
G
G
Jirillo
E
E
Papa
S
S
D000818:Animals; D002417:Cattle; D004024:Dicyclohexylcarbodiimide / Q000378:metabolism; D007700:Kinetics; D008081:Liposomes; D008929:Mitochondria, Heart / Q000201:enzymology*; D008970:Molecular Weight; D009840:Oligomycins / Q000494:pharmacology; D006180:Proton-Translocating ATPases / Q000302:isolation & purification / Q000378:metabolism*; D011522:Protons; D013367:Submitochondrial Particles / Q000201:enzymology*; D014357:Trypsin / Q000494:pharmacology
D008081:Liposomes; D009840:Oligomycins; D011522:Protons; D004024:Dicyclohexylcarbodiimide; D014357:Trypsin; D006180:Proton-Translocating ATPases