Chromatophore vesicles of Rhodobacter capsulatus contain on average one F(O)F(1)-ATP synthase each.
ATP synthase is a unique rotary machine that uses the transmembrane electrochemical potential difference of proton (Delta(H(+))) to synthesize ATP from ADP and inorganic phosphate. Charge translocation by the enzyme can be most conveniently followed in chromatophores of phototrophic bacteria (vesicles derived from invaginations of the cytoplasmic membrane). Excitation of chromatophores by a short flash of light generates a step of the proton-motive force, and the charge transfer, which is coupled to ATP synthesis, can be spectrophotometrically monitored by electrochromic absorption transients of intrinsic carotenoids in the coupling membrane. We assessed the average number of functional enzyme molecules per chromatophore vesicle. Kinetic analysis of the electrochromic transients plus/minus specific ATP synthase inhibitors (efrapeptin and venturicidin) showed that the extent of the enzyme-related proton transfer dropped as a function of the inhibitor concentration, whereas the time constant of the proton transfer changed only marginally. Statistical analysis of the kinetic data revealed that the average number of proton-conducting F(O)F(1)-molecules per chromatophore was approximately one. Thereby chromatophores of Rhodobacter capsulatus provide a system where the coupling of proton transfer to ATP synthesis can be studied in a single enzyme/single vesicle mode.
Feniouk
Boris A
BA
Division of Biophysics, Faculty of Biology/Chemistry, University of Osnabrück, D-49069 Osnabrück, Germany.
Cherepanov
Dmitry A
DA
Voskoboynikova
Natalia E
NE
Mulkidjanian
Armen Y
AY
Junge
Wolfgang
W
D000244:Adenosine Diphosphate / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D000900:Anti-Bacterial Agents / Q000737:chemistry; D020130:Bacterial Chromatophores / Q000737:chemistry*; D002455:Cell Division; D004563:Electrochemistry; D007477:Ions; D007700:Kinetics; D008564:Membrane Potentials; D010455:Peptides; D006180:Proton-Translocating ATPases / Q000737:chemistry* / Q000378:metabolism*; D011522:Protons; D016354:Rhodobacter capsulatus / Q000201:enzymology*; D013053:Spectrophotometry; D013997:Time Factors
D000900:Anti-Bacterial Agents; D007477:Ions; D010455:Peptides; D011522:Protons; C012953:efrapeptin; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D006180:Proton-Translocating ATPases