Excited-state electronic asymmetry of the special pair in photosynthetic reaction center mutants: absorption and Stark spectroscopy.

The electronic absorption line shape and Stark spectrum of the lowest energy Q(y)() transition of the special pair in bacterial reaction centers contain a wealth of information on mixing with charge transfer states and electronic asymmetry. Both vary greatly in mutants that perturb the chemical composition of the special pair, such as the heterodimer mutants, and in mutants that alter interactions between the special pair and the surrounding reaction center protein, such as those that add or remove hydrogen bonds. The conventional and higher-order Stark spectra of a series of mutants are presented with the aim of developing a systematic description of the electronic structure of the excited state of the special pair that initiates photosynthetic charge separation. The mutants L168HF, M197FH, L131LH and L131LH/M160LH/M197FH are known to have different hydrogen-bonding patterns to the special pair; however, they exhibit Stark effects that are very similar to wild type. By contrast, the addition of a hydrogen bond to the M-side keto carbonyl group of the special pair in M160LH greatly affects both the absorption and Stark spectra. The heterodimer special pairs, L173HL and M202HL, exhibit much larger Stark effects than wild type, with the greatest effect in the M-side mutant. Double mutants that combine the M-side heterodimer and a hydrogen-bond addition to the L-side of the special pair decrease the magnitude of the Stark effect. These results suggest that the electronic asymmetry of the dimer can be perturbed either by the formation of a heterodimer or by adding or deleting a hydrogen bond to a keto carbonyl group. From the pattern observed, it is concluded that the charge transfer state P(L)(+)P(M)(-) has a larger influence on the excited state of the dimer in wild type than the P(L)(-)P(M)(+)charge transfer state. Furthermore, asymmetry can be varied continuously, from extreme cases in which the heterodimer and hydrogen-bond effects work together, to cases in which hydrogen bonding offsets the effects of the heterodimer, to cases in which the homodimer is perturbed by hydrogen bonds. This leads to a unified model for understanding the effects of perturbations on the electronic symmetry of the special pair, and this can be connected with perturbations on the properties of many other systems such as donor-acceptor-substituted polyenes.