Spectroscopic studies of green-bacterial reaction center

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Electron transfer kinetics in the reaction center (Re) complex from the green sulfur bacterium Chlorobium tepidum which binds two cytochrome c (cyt c) were studied by flash-absorption spectroscopy. Photooxidized P84O^+ was reduced 95% and 80% after 1 ms by cyt c with t^<1/2> of about 65 mus and 100 muz s after the 1st and the 2nd flashes, respectively . The observed rates, the establishment of redox equilibrium after each flash and the total amount of photooxidizable cyt c are consistent with the presence of 2 equivalent cyt c per photooxidizable P840. The data were well fitted assuming a midpoint potential difference of -53 mV for electron transfer from one cyt c to P8404^+, * Em being independent of the oxidation state of the other cyt c. These observations support a model with 2 monoheme cyt c which are symmetrically arranged around the RC core. The purified RC complex binds 0.6-1.2 menaquinone-7 (MQ) . However, no transient signal due to MQ could be observed between 360 and 450 nm in the time window from 10 ns to 4 mu s. On the acceptor side, purified RC can stably hold up to 3 electrons in the mu s time range.However, after the 4th flash, a charge recombination reaction with t^<1/2> of 20 ns was first observed. From our data, there is no evidence for involvement of MQ in charge separation in the RC of green sulfur bacteria.