The local electric field within phospholipid membranes modulates the charge transfer reactions in reaction centres.
Three different cholesterol derivatives and phloretin, known to affect the local electric field in phospholipid membranes, have been introduced into Rhodobacter sphaeroides reaction centre-containing phospholipid liposomes. We show that cholesterol and 6-ketocholestanol significantly slow down the interquinone first electron transfer (approximately 10 times), whereas phloretin and 5-cholesten-3beta-ol-7-one leave the kinetics essentially unchanged. Interestingly, the two former compounds have been shown to increase the dipole potential, whereas the two latter decrease it. We also measured in isolated RCs the rates of the electron and proton transfers at the first flash. Over the pH range 7-10.5 both reactions display biphasic behaviors with nearly superimposable rates and amplitudes, suggesting that the gating process limiting the first electron transfer is indeed the coupled proton entry. We therefore interpret the effects of cholesterol and 6-ketocholestanol as due to dipole concentration producing an increased free energy barrier for protons to enter the protein perpendicular to the membrane. We also report for the first time in R. sphaeroides RCs, at room temperature, a biphasicity of the P(+)Q(A)(-) charge recombination, induced by the presence of cholesterol derivatives in proteoliposomes. We propose that these molecules decrease the equilibration time between two RC conformations, therefore revealing their presence.
Pilotelle-Bunner
Anne
A
Laboratoire de Chimie Physique, UMR 8000, Université Paris-Sud 11/CNRS, Orsay, France.
Beaunier
Patricia
P
Tandori
Julia
J
Maroti
Peter
P
Clarke
Ronald J
RJ
Sebban
Pierre
P
D000880:Anthraquinones / Q000737:chemistry; D002462:Cell Membrane / Q000737:chemistry; D002784:Cholesterol / Q000031:analogs & derivatives / Q000737:chemistry; D004574:Electromagnetic Fields; D004579:Electron Transport; D006863:Hydrogen-Ion Concentration; D007653:Ketocholesterols / Q000737:chemistry; D007700:Kinetics; D008081:Liposomes / Q000737:chemistry; D046529:Microscopy, Electron, Transmission; D008958:Models, Molecular; D010693:Phloretin / Q000737:chemistry; D010713:Phosphatidylcholines / Q000737:chemistry; D010743:Phospholipids / Q000737:chemistry*; D045322:Photosynthetic Reaction Center Complex Proteins / Q000737:chemistry*; D012242:Rhodobacter sphaeroides / Q000737:chemistry; D013696:Temperature; D013816:Thermodynamics
D000880:Anthraquinones; D007653:Ketocholesterols; D008081:Liposomes; D010713:Phosphatidylcholines; D010743:Phospholipids; D045322:Photosynthetic Reaction Center Complex Proteins; C063833:6-ketocholestanol; D002784:Cholesterol; C003001:7-ketocholesterol; D010693:Phloretin