Charge Separation in Rhodobacter sphaeroides Mutant Reaction Centers with Increased Midpoint Potential of the Primary Electron Donor

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A. Yu. Khmelnitskiy^*, R. A. Khatypov, A. M. Khristin, M. M. Leonova, L. G. Vasilieva, and V. A. Shuvalov

Institute of Basic Biological Problems, Russian Academy of Sciences, 142290 Pushchino, Moscow Region, Russia; fax: (496) 773-0532; E-mail: hmelan@gmail.com

^* To whom correspondence should be addressed.

Received August 8, 2012; Revision received September 4, 2012

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Primary charge separation dynamics in four mutant reaction centers (RCs) of the purple bacterium Rhodobacter sphaeroides with increased midpoint potential of the primary electron donor P (M160LH, L131LH, M197FH, and M160LH + L131LH + M197FH) have been studied by femtosecond transient absorption spectroscopy at room temperature. The decay of the excited singlet state in the wild-type and mutant RCs is complex and has two main exponential components, which indicates heterogeneity of electron transfer rates or the presence of reverse electron transfer reactions. The radical anion band of monomeric bacteriochlorophyll B_A at 1020 nm was first observed in transient absorbance difference spectra of single mutants. This band remains visible, although with somewhat reduced amplitude, even at delays up to tens of picoseconds when stimulated emission is absent and the reaction centers are in the P⁺H_A^– state. The presence of this band in this time period indicates the existence of thermodynamic equilibrium between the P⁺B_A^–H_A and P⁺B_AH_A^– states. The data give grounds for assuming that the value of the energy difference between the states P*, P⁺B_A^–H_A, and P⁺B_AH_A^– at early times is of the same order of magnitude as the energy kT at room temperature. Besides, monomeric bacteriochlorophyll B_A is found to be an immediate electron acceptor in the single mutant RCs, where electron transfer is hampered due to increased energy of the P⁺B_A^– state with respect to P*.

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KEY WORDS: photosynthesis, charge separation, reaction center, femtosecond spectroscopy, electron transfer

DOI: 10.1134/S0006297913010070

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