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Long-lived primary radical pair state detected by time-resolved fluorescence and absorption spectroscopy in an isolated Photosystem two core

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Abstract

A Photosystem two (PS II) core preparation containing the chlorophyll a binding proteins CP 47, CP 43, D1 and D2, and the non-chlorophyll binding cytochrome-b559 and 33 kDA polypeptides, has been isolated from PS II-enriched membranes of peas using the non-ionic detergent heptylthioglucopyranoside and elevated ionic strengths. The primary radical pair state, P680+Pheo-, was studied by time-resolved absorption and fluorescence spectroscopy, under conditions where quinone reduction and water-splitting activities were inhibited. Charge recombination of the primary radical pair in PS II cores was found to have lifetimes of 17.5 ns measured by fluorescence and 21 ns measured by transient decay kinetics under anaerobic conditions. Transient absorption spectroscopy demonstrated that the activity of the particles, based on primary radical pair formation, was in excess of 70% (depending on the choice of kinetic model), while time-resolved fluorescence spectroscopy indicated that the particles were 91% active. These estimates of activity were further supported by steady-state measurements which quantified the amount of photoreducible pheophytin. It is concluded that the PS II core preparation we have isolated is ideal for studying primary radical pair formation and recombination as demonstrated by the correlation of our absorption and fluorescence transient data, which is the first of its kind to be reported in the literature for isolated PS II core complexes from higher plants.

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Abbreviations

CP 43 and CP 47:

chlorophyll binding proteins of PS II having apparent molecular weights on SDS-PAGE of 43 kDa and 47 kDa, respectively

D1 and D2 polypeptides:

PS II reaction centre polypeptides encoded by the psbA and psbD genes, respectively

HPLC:

high performance liquid chromatography

PS II:

Photosystem two

SDS-PAGE:

sodium dodecyl sulphate-polyacrylamide gel electrophoresis

P680:

primary electron donor of PS II

Pheo:

phenophytin a

SPC:

single photon counting

PBQ:

phenyl-p-benzoquinone

DPC:

1,5-diphenylcarbazide

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Author notes

  1. P. J. Booth

    Present address: Section de Bioenergetique, Departement de Biologie Cellulaire et Moleculaire, Centre d'Etudes Nucléaires de Saclay, 91191, Gif-sur-Yvette, Cedex, France

Authors and Affiliations

  1. Photochemistry Research Group, Department of Biology, Imperial College of Science, Technology & Medicine, London

    B. Crystall, J. R. Durrant, S. Özer, G. Porter & D. R. Klug

  2. Physical Chemistry Laboratory, Oxford University, Oxford, UK

    P. J. Booth

Authors
  1. J. De Las Rivas
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  2. B. Crystall
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  3. P. J. Booth
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  4. J. R. Durrant
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  5. S. Özer
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  6. G. Porter
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  7. D. R. Klug
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  8. J. Barber
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AFRC Photosynthesis Research Group, Department of Biochemistry

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De Las Rivas, J., Crystall, B., Booth, P.J. et al. Long-lived primary radical pair state detected by time-resolved fluorescence and absorption spectroscopy in an isolated Photosystem two core. Photosynth Res 34, 419–431 (1992). https://doi.org/10.1007/BF00029816

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