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Kinetics and yields of bacteriochlorophyll fluorescence: redox and conformation changes in reaction center of Rhodobacter sphaeroides

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Abstract

Induction of the bacteriochlorophyll fluorescence under rectangular shape of intense laser diode illumination (1 W cm−2, 808 nm) was measured over wide time range from 10 μs to 4 s in whole cells, chromatophore and isolated reaction center protein of wild type and carotenoid-less mutant (R-26.1) of purple photosynthetic bacterium Rhodobacter sphaeroides. While the antenna-containing species showed large and positive variable fluorescence (F v) to initial fluorescence (F 0) (F v/F 0 ∼ 4.5 in whole cells), the isolated RC had negative change (F v/F 0 ∼ −0.6) during photochemistry. In chromatophore from R-26.1, only seven times higher rate was measured than in isolated reaction center under identical experimental conditions. The enhancement effect of large antenna on the rate of photochemistry in chromatophore was partially compensated by the favorable pigment absorption properties in isolated RC. The transition from membrane bound to isolated form of the reaction center was probed by titration of zwitterionic detergent LDAO in chromatophore, and at 0.03% LDAO concentration, sharp change of the variable fluorescence was observed. The sudden drop was explained by the formation of LDAO micelles. After the photochemical phase, additional change of fluorescence yield could be observed in isolated RC considered as manifestation of long-living conformations of the trapped redox states of the protein characterized by non-exponential kinetics. Strong support was provided for use of the fluorescence induction to track structural and conformation changes at their earliest phases in chromatophores and isolated reaction centers.

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Abbreviations

BChl:

Bacteriochlorophyll

Bpheo:

Bacteriopheophytin

D:

External electron donor to P+

F 0 :

Initial fluorescence intensity

F v :

Variable fluorescence intensity

LDAO:

N,N-dimethyldodecylamine-N-oxide

LH I and LH II:

Light harvesting complexes I and II, respectively

P:

Bacteriochlorophyll dimer

PSII:

Photosystem II of higher plants

Q:

Quinone acceptor

QA :

Primary quinone acceptor

QB :

Secondary quinone acceptor

RC:

Reaction center protein

UQ10 :

Ubiquinone-50

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Acknowledgments

This work was supported by NKTH-OTKA (K-67850), MTA-CNR and TéT. Thanks are due to Dr. L. Gerencsér and Ms. R. Kékesi for their help in the simulation and preliminary measurements.

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  1. Department of Biophysics, University of Szeged, Egyetem utca 2, 6723, Szeged, Hungary

    Péter Maróti

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  1. Péter Maróti
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Correspondence to Péter Maróti.

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Regional Biophysics Conference of the National Biophysical Societies of Austria, Croatia, Hungary, Italy, Serbia, and Slovenia.

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Maróti, P. Kinetics and yields of bacteriochlorophyll fluorescence: redox and conformation changes in reaction center of Rhodobacter sphaeroides . Eur Biophys J 37, 1175–1184 (2008). https://doi.org/10.1007/s00249-008-0300-5

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