Stoichiometrically bound β-carotene in the cytochrome b6f complex of oxygenic photosynthesis protects against oxygen damage

Abstract

The cytochrome b6f complex of oxygenic photosynthesis carries out 'dark reactions' of electron transfer that link the light-driven reactions of the reaction centers, and coupled proton transfer that generates part of the electrochemical potential utilized for ATP synthesis. In contrast to the bc1 complex of the respiratory chain, with which there are many structural and functional homologies, the b6f complex contains bound pigment molecules. Along with the specifically bound chlorophyll a previously found to be bound stoichiometrically in the dimeric b6f complex, it was found in the present study that β-carotene is also present in the b6f complex at stoichiometric levels or nearly so. Chlorophyll and carotenoid pigments were quantitatively extracted from b6f complex purified from (i) the thermophilic cyanobacterium, Mastigocladus laminosus, (ii) spinach chloroplasts, and (iii) the green alga, Chlamydomonas reinhardtii. Visible and mass spectra showed the carotenoid to be a β-carotene of molecular weight = 536, with a stoichiometry of 1.0:1 relative to cytochrome f in the highly active M. laminosus complex but somewhat lower stoichiometries, 0.77 and 0.55, in the b6f complex obtained from spinach chloroplasts and C. reinhardtii. A photoprotective function for the β-carotene was inferred from the findings that the rate of photobleaching of the chlorophyll a bound in the complex was found to vary inversely with β-carotene content and to decrease markedly in the presence of ambient N2 instead of air. The presence of β-carotene in the b6f complex, and not in the related bc1 complexes of the mitochondrial respiratory chain and photosynthetic bacteria, suggests that an additional function is to protect the protein complexes in oxygenic photosynthetic membranes against toxic effects of intramembrane singlet O2.