Spectroscopic studies on electron transfer between plastocyanin and cytochrome b6f complex

This paper reports the results of the research on the interaction between the highly active cytochrome b6f complex and plastocyanin, both isolated from the same source – spinachia oleracea plants. An equilibrium constant K between the cytochrome f of the cytochrome b6f complex and plastocyanin has been estimated by two independent spectroscopic techniques: steady-state absorption spectroscopy and stopped-flow. The second-order rate constants k2 for forward and backward electron transfer between cytochrome f and plastocyanin have been found between 1.4–2 × 10^7 and 8–10 × 10^6 M^-1 s^-1, respectively, giving the value of an equilibrium constant of about 2 (plus-minus) 0.4 or a difference in redox potential between plastocyanin and cytochrome f of cytochrome b6f complex of ca. 17 mV. The value of K=1.7(plus-minus)0.3 has been estimated from steady-state experiments in which the initial and final concentrations of participating components after mixing have been estimated via differential spectra analysis or spectra deconvolution. We propose a method of evaluation of the final plastocyanin concentration after the electron transfer reaction between cytochrome bf complex and plastocyanin that overcomes the interference by the strong chlorophyll absorption in the spectral region where oxidised plastocyanin has its low extinction absorption band. The data from both experiments, in the system devoid of quinol being the electron donor to cytochrome b6, suggest that in case of electron transfer from cytochrome f to plastocyanin electron transfer can either bypass cytochrome f or the Rieske iron–sulfur protein can be reduced prior to its movement to the quinol binding site of cytochrome b6. The role of the Rieske protein in forward and backward electron transfer reactions is discussed.