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Charge separation in Rhodobacter sphaeroides mutant reaction centers with increased midpoint potential of the primary electron donor

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Abstract

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 BA 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 HA and P+BAH −A states. The data give grounds for assuming that the value of the energy difference between the states P*, P+B −A HA, and P+BAH −A at early times is of the same order of magnitude as the energy kT at room temperature. Besides, monomeric bacteriochlorophyll BA 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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Abbreviations

ΔA :

absorption change (light-minus-dark)

BA and BB :

monomeric BChl in A- and B-chain, respectively

BChl:

bacteriochlorophyll

BPheo:

bacteriopheophytin

HA and HB :

BPheo in A- and B-chain, respectively

P:

primary electron donor, BChl dimer

PA and PB :

BChl molecules within P

QA and QB :

primary and secondary quinone, respectively

Rba., Rhodobacter :

RC, reaction center

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Authors and Affiliations

  1. Institute of Basic Biological Problems, Russian Academy of Sciences, 142290, Pushchino, Moscow Region, Russia

    A. Yu. Khmelnitskiy, R. A. Khatypov, A. M. Khristin, M. M. Leonova, L. G. Vasilieva & V. A. Shuvalov

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  1. A. Yu. Khmelnitskiy
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  2. R. A. Khatypov
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  3. A. M. Khristin
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  4. M. M. Leonova
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  5. L. G. Vasilieva
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  6. V. A. Shuvalov
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Correspondence to A. Yu. Khmelnitskiy.

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Published in Russian in Biokhimiya, 2013, Vol. 78, No. 1, pp. 82–91.

Originally published in Biochemistry (Moscow) On-Line Papers in Press, as Manuscript BM12-226, December 9, 2012.

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Khmelnitskiy, A.Y., Khatypov, R.A., Khristin, A.M. et al. Charge separation in Rhodobacter sphaeroides mutant reaction centers with increased midpoint potential of the primary electron donor. Biochemistry Moscow 78, 60–67 (2013). https://doi.org/10.1134/S0006297913010070

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  • DOI: https://doi.org/10.1134/S0006297913010070

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