Temperature dependence of charge recombination from states D+QA- and D+QB- in photosynthetic reaction centers isolated from the thermophilic bacterium Chloroflexus aurantiacus

The temperature dependence of charge recombination from the P+Q(A)- and from the P+Q(B)- states produced by a flash was studied in reaction centers isolated from the photosynthetic thermophilic bacterium Chloroflexus aurantiacus. P designates the primary electron donor; Q(A) and Q(B) the primary and secondary quinone electron acceptors respectively. In Q(B)-depleted reaction centers the rate constant (k(AP)) for P+Q(A)- recombination was temperature independent between 0-50°C (17.6 ± 0.7 s-1 at pH 8 and pH 10). The same value was obtained in intact membranes in the presence of o-phenanthroline. Upon lowering the temperature from 250 K to 160 K, k(AP) increased by a factor of two and remained constant down to 80K. The overall temperature dependence of k(AP) was consistent with an activationless process. Ubiquinone (UQ-3) and different types of menaquinone were used for Q(B) reconstitution. In UQ-3-reconstituted reaction centers charge recombination was monoexponential (rate constant k = 0.18 ± 0.03 s-1) and temperature independent between 5-40°C. In contrast, in menaquinone-3- and menaquinone-4-reconstituted reaction centers P+ rereduction following a flash was markedly biphasic and temperature dependent. In menaquinone-6-reconstituted reaction centers a minor contribution from a third kinetic phase corresponding to P+Q(A)- charge recombination was detected. Analysis of these kinetics and of the effects of the inhibitor o-phenanthroline at high temperature suggest that in detergent suspensions of menaquinone-reconstituted reaction centers a redox reaction removing electrons from the quinone acceptor complex competes with charge recombination. Instability of the semiquinone anions is more pronounced when Q(B) is a short-chain menaquinone. From the temperature dependence of P+ decay the activation parameters for the P+Q(B)- recombination and for the competing side oxidation of the reduced menaquinone acceptor have been derived. For both reactions the activation enthalpies and entropies change markedly with menaquinone chain length but counterbalance each other, resulting in activation free energies at ambient temperature independent of the menaquinone tail. When reaction centers are incorporated into phospholipid vesicles containing menaquinone-8 a temperature-dependent, monophasic, o-phenanthroline-sensitive recombination from the P+Q(B)- state is observed, which is consistent with the formation of stable semiquinone anions. This result seems to indicate a proper Q(B) functioning in the two-subunit reaction center isolated from Chloroflexus aurantiacus when the complex is inserted into a lipid bilayer.