Manipulating the direction of electron transfer in the bacterial reaction center by swapping Phe for Tyr near BChlM (L181) and Tyr for Phe near BChlL (M208)

We have investigated the primary charge separation processes in Rb. capsulatus reaction centers (RCs) bearing the mutations Phe(L181) --> Tyr, Tyr(M208) --> Phe, and Leu(M212) --> His. In the YFH mutant, decay of the excited primary electron donor P* occurs with an 11 +/- 2 ps time constant and is trifurcated to give (1) internal conversion to the ground state (similar to 10% yield), (2) charge separation to the L side of the RC (similar to 60% yield). and (3) electron transfer to the M-side bacteriopheophytin BPhM (similar to 30% yield). These results relate previous work in which the ionizable residues Lys (at L178) and Asp (at M201) have been used to facilitate charge separation to the M side of the RC, and the widely studied L181 and M208 mutants. One conclusion that comes from this work is that the Tyr (M208) --> Phe and Gly(M201) --> Asp mutations near the L-side bacteriochlorophyll (BChl(L)) raise the free energy of P(+)BChlL(-) by comparable amounts. The results also suggest that the free energy of P(+)BChl(M)(-) is lowered more substantially by a Tyr at L181 than a Lys at L178. The results on the YFH mutant further demonstrate that the free energy differences between the L- and M-side charge-separated states play a significant role in the directionality of charge separation in the wild-type RC. and place limits on the contributing role of differential electronic matrix elements on the two sides of the RC.