Two Pathways to Understanding Electron Transfer in Reaction Centers from Photosynthetic Bacteria: A Comparison of Rhodobacter Sphaeroides and Rhodobacter Capsulatus Mutants

The rates, yields, mechanisms and directionality of electron transfer (ET) are explored in twelve pairs of Rhodobacter (R.) sphaeroides and R. capsulatus mutant RCs designed to defeat ET from the excited primary donor (P*) to the A-side cofactors and re-direct ET to the normally inactive mirror-image B-side cofactors. In general, the R. sphaeroides variants have larger P+H-B(-) yields (up to similar to 90%) than their R. capsulatus analogs (up to similar to 60%), where HB is the B-side bacteriopheophytin. Substitution of Tyr for Phe at L-polypeptide position L181 near BB primarily increases the contribution of fast P*. P+BB- -> P+HB- two-step ET, where B-B is the "bridging" B-side bacteriochlorophyll. The second step (similar to 6-8 ps) is slower than the first (similar to 3-4 ps), unlike A-side two-step ET (P* -> P+BA- -> P+HA-) where the second step (similar to 1 ps) is faster than the first (similar to 3-4 ps) in the native RC. Substitutions near HB, at L185 (Leu, Trp or Arg) and at M-polypeptide site M133/131 (Thr, Val or Glu), strongly affect the contribution of slower (20-50 ps) P* -> P+HB- one-step superexchange ET. Both ET mechanisms are effective in directing electrons "the wrong way" to HB and both compete with internal conversion of P* to the ground state (similar to 200 ps) and ET to the A-side cofactors. Collectively, the work demonstrates cooperative amino-acid control of rates, yields and mechanisms of ET in bacterial RCs and how A- vs. B-side charge separation can be tuned in both species.