DCCD-sensitive proton permeability of bacterial photosynthetic membranes. Cross-reconstitution studies with purified bovine heart Fo subunits.
The DCCD-sensitive proton permeability of chromatophores, from a green strain of Rhodobacter Capsulatus is potentiometrically detected following the proton release induced by a transmembrane diffusion potential imposed by a valinomycin-mediated potassium influx with a procedure already used for bovine heart submitochondrial particles (ESMP) and vesicles from Escherichia coli (Zanotti et al. (1994) Eur. J. Biochem. 222, 733-741). In the photosynthetic system, addition of increasing amounts of DCCD inhibits, with a similar titre, both proton permeability and MgATP-dependent ATPase activity as detected in the dark. The titre for 50% inhibition coincides with that obtained measuring proton permeability and ATP hydrolysis in ESMP. Upon removal of F1, the passive proton permeability is much less sensitive to DCCD in chromatophores than in USMP, suggesting that in chromatophores the F1-Fo interaction shapes the DCCD-sensitive proton conducting pathway. Addition of the purified mitochondrial FoI-PVP and oligomycin sensitivity-conferring (OSCP) proteins to the F1 stripped chromatophores restored the sensitivity of proton permeability to DCCD detected in untreated chromatophores. Analysis of the binding of 14C[DCCD] on F1 stripped chromatophores shows that the increase of DCCD sensitivity of proton permeability, caused by addition of mitochondrial Fo proteins, is related to an increase of the binding of the inhibitor to subunit c of Fo sector of ATP synthase complex.
Zanotti
F
F
Institute of Medical Biochemistry and Chemistry, University of Bari, Italy.
Casadio
R
R
Perrucci
C
C
Guerrieri
F
F
D000818:Animals; D020130:Bacterial Chromatophores / Q000187:drug effects* / Q000378:metabolism*; D002417:Cattle; D004024:Dicyclohexylcarbodiimide / Q000378:metabolism / Q000494:pharmacology*; D066298:In Vitro Techniques; D007700:Kinetics; D008929:Mitochondria, Heart / Q000201:enzymology; D010539:Permeability; D010788:Photosynthesis; D011487:Protein Conformation; D006180:Proton-Translocating ATPases / Q000737:chemistry / Q000378:metabolism*; D011522:Protons; D016354:Rhodobacter capsulatus / Q000187:drug effects* / Q000378:metabolism*
D011522:Protons; D004024:Dicyclohexylcarbodiimide; D006180:Proton-Translocating ATPases