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pH sensitivity of the redox state of cytochrome b559 may regulate its function as a protectant against donor and acceptor side photoinhibition

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  • Oxygenic Photosynthesis
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Abstract

A series of experiments have been conducted with isolated reaction centers of photosystem two (PS II) with the aim to elucidate the functional role of cytochrome (Cyt b 559). At pH 6.5 it was found that Cyt b 559 was reversibly photoreduced by red actinic light when Mn2+ was present as an electron donor while at pH 8.5 a photo-oxidation was observed under the same lighting conditions, which was dark reversible in the presence of hydroquinone. These pH dependent light induced changes were measured under anaerobic conditions and correlated with changes in the relative levels of high (HP) and low (LP) potential forms of the cytochrome. At pH 6.5 the cytochrome was mainly in its LP form while at pH 8.5 a significant proportion was converted to the HP form as detected by dark titrations with hydroquinone. This pH dependent difference in the levels of HP and LP Cyt b 559 was also detected when bright white light was used to monitor the level of the LP form using a novel reaction involving direct electron donation from the flavin of glucose oxidase (present in the medium and used together with glucose and catalase as an oxygen trap). The results suggest that PS II directly oxidises and reduces the HP and LP forms, respectively and that the extent of these photo-reactions is dependent on the relative levels of the two forms, which are in turn governed by the pH. This conclusion is interpreted in terms of the model presented previously (Barber J and De Las Rivas J (1993) Proc Natl Acad Sci USA 90: 10942–10946) whereby the pH induced effect is considered as a possible mechanism by which interconversion of LP and HP forms of Cyt b 559 is achieved. In agreement with this was the finding that as the extent of photo-oxidisable HPCyt b 559 increases, with increasing pH, the rate of irreversible photo-oxidation of β-carotene decreases, a result expected if the HP form protects against donor side photoinhibition.

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Abbreviations

β-car:

β-carotene

CCCP:

carbonylcyanide m-chloro-phenylhydrazone

Chl:

chlorophyll

Cyt b 559 :

cytochrome b 559

HPCyt b 559 :

high potential form of cytochrome b 559 which is reducible by hydroquinone

LPCyt b 559 :

low potential form of cytochrome b 559 which is non-reducible by hydroquinone

D1 and D2:

products of the psbA and psbD genes, respectively

LHC II:

light-harvesting chlorophyll protein complex associated with PS II

Mes:

2-(N-morpholino) ethanesulphonic acid

P680:

primary electron donor of PS II

Pheo:

pheophytin

PQ:

plastoquinone

PS II:

Photosystem II

QA :

first stable quinone electron acceptor of PS II

QB :

second stable quinone electron acceptor of PS II

RC:

reaction center

SDS:

sodium dodecyl sulphate

SiMo:

silicomolybdate

Tris:

tris(hydroxymethyl) amino methane

YZ and YD :

tyrosine residues 161 in D1 and D2 proteins of the PS II RC which act as secondary electron donors to P680

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Author notes

  1. Jayier De Las Rivas

    Present address: Department of Biochemistry and Molecular Biology, Faculty of Sciences, University of the Basque Country, P.O. Box 644, Bilbao, Spain

  2. Judith Klein

    Present address: Department of Genetics, University of Cologne, 5000, Cologne 41, Germany

Authors and Affiliations

  1. Photosynthesis Research Group, Wolfson Laboratories, Department of Biochemistry, Imperial College of Science, Technology & Medicine, London, SW7 2AY, UK

    Jayier De Las Rivas, Judith Klein & James Barber

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  1. Jayier De Las Rivas
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  2. Judith Klein
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De Las Rivas, J., Klein, J. & Barber, J. pH sensitivity of the redox state of cytochrome b559 may regulate its function as a protectant against donor and acceptor side photoinhibition. Photosynth Res 46, 193–202 (1995). https://doi.org/10.1007/BF00020430

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  • DOI: https://doi.org/10.1007/BF00020430

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