The binding of quinone to the photosynthetic reaction centers: kinetics and thermodynamics of reactions occurring at the QB-site in zwitterionic and anionic liposomes.
Liposomes represent a versatile biomimetic environment for studying the interaction between integral membrane proteins and hydrophobic ligands. In this paper, the quinone binding to the QB-site of the photosynthetic reaction centers (RC) from Rhodobacter sphaeroides has been investigated in liposomes prepared with either the zwitterionic phosphatidylcholine (PC) or the negatively charged phosphatidylglycerol (PG) to highlight the role of the different phospholipid polar heads. Quinone binding (K Q) and interquinone electron transfer (L AB) equilibrium constants in the two type of liposomes were obtained by charge recombination reaction of QB-depleted RC in the presence of increasing amounts of ubiquinone-10 over the temperature interval 6-35 °C. The kinetic of the charge recombination reactions has been fitted by numerically solving the ordinary differential equations set associated with a detailed kinetic scheme involving electron transfer reactions coupled with quinone release and uptake. The entire set of traces at each temperature was accurately fitted using the sole quinone release constants (both in a neutral and a charge separated state) as adjustable parameters. The temperature dependence of the quinone exchange rate at the QB-site was, hence, obtained. It was found that the quinone exchange regime was always fast for PC while it switched from slow to fast in PG as the temperature rose above 20 °C. A new method was introduced in this paper for the evaluation of constant K Q using the area underneath the charge recombination traces as the indicator of the amount of quinone bound to the QB-site.
Department of Chemistry, University of Bari, 70126, Bari, Italy.
Fabio
F
Mavelli
Massimo
M
Trotta
Fulvio
F
Ciriaco
Angela
A
Agostiano
Livia
L
Giotta
Francesca
F
Italiano
Francesco
F
Milano
D007700:Kinetics; D008081:Liposomes / Q000737:chemistry* / Q000378:metabolism*; D008954:Models, Biological; D010713:Phosphatidylcholines / Q000378:metabolism; D010715:Phosphatidylglycerols / Q000378:metabolism; D045322:Photosynthetic Reaction Center Complex Proteins / Q000378:metabolism*; D011485:Protein Binding; D011809:Quinones / Q000378:metabolism*; D012242:Rhodobacter sphaeroides / Q000201:enzymology; D013696:Temperature; D013816:Thermodynamics
D008081:Liposomes; D010713:Phosphatidylcholines; D010715:Phosphatidylglycerols; D045322:Photosynthetic Reaction Center Complex Proteins; D011809:Quinones