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High-light-induced superoxide anion radical formation in cytochrome b 6 f complex from spinach as detected by EPR spectroscopy

  • Original Papers
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Photosynthetica

An Erratum to this article was published on 01 September 2011

Abstract

The generation of superoxide anion radical (O2 ·−) in the cytochrome b 6 f complex (Cyt b 6 f) of spinach under high-light illumination was studied using electron paramagnetic resonance spectroscopy. The generation of O2 ·− was lost in the absence of molecular oxygen. It was also suppressed in the presence of NaN3 and could be scavenged by extraneous antioxidants such as ascorbate, β-carotene, and glutathione. The results also indicate that O2 ·−, which is produced under high-light illumination of the Cyt b 6 f from spinach, might be generated from a reaction involing 1O2, and the Rieske Fe-S protein could serve as the electron donor in the O2 ·− production. The mechanism of photoprotection of the Cyt b 6 f complex by antioxidants is discussed.

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Abbreviations

Car:

carotene

Chl a :

chlorophyll a

3Chl a*:

triplet excited state of chlorophyll a

Cyt b 6 f :

cytochrome b 6 f complex

EMPO:

2-ethoxycarbonyl-2-methyl-3,4-dihydro-2H-pyrrole-1-oxide

EPR:

electron paramagnetic resonance

O2 ·− :

superoxide anion radical

ROS:

reactive oxygen species

1O2:

singlet oxygen

TEMP:

2,2,6,6-tetramethylpiperidine

β-DM:

n-dodecyl-β-D-maltoside

β-OG:

n-octyl-β-D-glucopyranoside

References

  • Allen, J.F.: Cytochrome b 6 f: structure for signaling and vectorial metabolism. — Trends Plant Sci. 9: 130–137, 2004.

    Article  PubMed  CAS  Google Scholar 

  • Asada, K., Kiso, K., Yoshikawa, K.: Univalent reduction of molecular oxygen by spinach chloroplasts on illumination. — J. Biol. Chem. 249: 2175–2181, 1974.

    PubMed  CAS  Google Scholar 

  • Asada, K.: Production and scavenging of reactive oxygen species in chloroplasts and their functions. — Plant. Physiol. 141: 391–396, 2006.

    Article  PubMed  CAS  Google Scholar 

  • Buettner, G.R., Oberley, L.W.: The apparent production of superoxide and hydroxyl radicals by hematoporphyrin and light as seen by spin-trapping. — FEBS Letters 121: 161–164, 1980.

    Article  PubMed  CAS  Google Scholar 

  • Chen, X.B., Zhao, X.H., Zhu, Y., Gong, Y.D., Li, L.B., Zhang, J.P., Kuang, T.Y.: Hydrogen peroxide-induced chlorophyll a bleaching in the cytochrome b 6 f complex: a simple and effective assay for stability of the complex in detergent solutions. — Photosyhth. Res. 90: 205–214, 2006.

    Article  CAS  Google Scholar 

  • Cramer, W.A., Zhang, H.M., Yan, J.S., Kurisu, G., Smith, J.L. Transmembrane traffic in the cytochrome b 6 f complex. — Annu. Rev. Biochem. 75: 769–90, 2006.

    Article  PubMed  CAS  Google Scholar 

  • Crystall, B., Booth, P.J., Klug, D.R., Barber, J., Porter, G.: Resolution of a long lived fluorescence component from D1/D2 cytochrome b-559 reaction centres. — FEBS Lett. 249: 75–78, 1989.

    Article  CAS  Google Scholar 

  • Doyle, M.P., Li, L.B., Yu, L., Yu, C.A.: Identification of a Mr = 17,000 protein as the plastoquinone-binding protein in the Cytochrome b 6 f complex from spinach chloroplasts. — J. Biol. Chem. 264:1387–92, 1989.

    PubMed  CAS  Google Scholar 

  • Edreva, A.: Generation and scavenging of reactive oxygen species in chloroplasts: a submolecular approach. — Agr. Ecosyst. Environ. 106: 119–133, 2005.

    Article  CAS  Google Scholar 

  • Furbank, R.T., Badger, M.R., Osmond, C.B.: Photoreduction of oxygen in mesophyll chloroplasts of C4 plants. A model system for studying an in vivo Mehler reaction — Plant Physiol. 73: 1038–1041, 1983.

    Article  PubMed  CAS  Google Scholar 

  • Genova, M.L., Ventura, B., Giuliano, G., Bovina, C., Formiggini, G., Castelli, G.P., Lenaz, G.: The site of production of superoxide radical in mitochondrial Complex I is not a bound ubisemiquinone but presumably iron-sulfur cluster N2. — FEBS Lett. 505: 364–368, 2001.

    Article  PubMed  CAS  Google Scholar 

  • Hurt, E., Hauska, G., Malkin, R.: Isolation of the Rieske ironsulfur protein from the cytochrome b 6/f complex of spinach chloroplasts. — FEBS Lett. 134: 1–5, 1981.

    Article  PubMed  CAS  Google Scholar 

  • Krinsky, N.I.: Biological roles of singlet oxygen. — In: H.H. Wasserman, (ed.): Singlet Oxygen. Vol. 40, Pp. 597–641. Academic Press, New York 1979.

    Google Scholar 

  • Kurisu, G., Zhang, H.M., Smith, J.L., Cramer, W.A.: Structure of the cytochrome b 6 f complex of oxygenic photosynthesis: Tuning the cavity. — Science 302: 1009–1014, 2003.

    Article  PubMed  CAS  Google Scholar 

  • Li, L.B., Zou, Y.P., Yu, L., Yu, C.A.: The catalytic role of subunit IV of the Cytochrome b 6-f complex from spinach chloroplasts. — Biochim. Biophys. Acta 1057: 215–222, 1991.

    Article  PubMed  CAS  Google Scholar 

  • Lion, Y., Delmelle, M., van de Vorst, A.: New method of detecting singlet oxygen production. — Nature 263: 442–443, 1976.

    Article  PubMed  CAS  Google Scholar 

  • Lion, Y., Gandin, E., van de Vorst, A.: On the production of nitroxide radicals by singlet oxygen reaction: An electronparamagneticres study. — Photochem. Photobiol. 31: 305–309, 1980.

    Article  CAS  Google Scholar 

  • Mehler, A.D.: Studies on the reactions of illuminated chloroplasts. II. Stimulation on inhibition of the reaction with molecular oxygen. — Arch. Biochem. Biophys. 34: 339–351, 1951.

    Article  PubMed  CAS  Google Scholar 

  • Mitchell, P.: Chemiosmotic coupling in oxidative and photosynthetic phosphorylation. — Biol. Rev. Cambridge Phil. Soc. 41: 445–602, 1966.

    Article  CAS  Google Scholar 

  • Olive, G., Mercier, A., Le Moigne, F., Rockenbauer, A., Tordo, P.: 2-ethoxycarbonyl-2-methyl-3,4-dihydro-2H-pyrrole-1-oxide: evaluation of the spin trapping properties. — Free Radical. Biol. Med. 28: 403–408, 2000.

    Article  CAS  Google Scholar 

  • Pospíšil, P., Arató, A., Liszkay, A.K., Rutherford, W.A.: Hydroxyl radical generation by photosystem II. — Biochemistry 43: 6783–6792, 2004.

    Article  PubMed  Google Scholar 

  • Pospíšil, P., Šnyrychová, I., Kruk, J., Strźalka, K., Nauš, J.: Evidence that cytochrome b559 is involved in superoxide production in photosystem II: effect of synthetic shortchain plastoquinones in a cytochrome b559 tobacco mutant. — Biochem. J. 397: 321–327, 2006.

    Article  PubMed  Google Scholar 

  • Pospíšil, P.: Production of reactive oxygen species by photosystem II. — Biochim. Biophys. Acta-B 1787: 1151–1160, 2009.

    Article  Google Scholar 

  • Pierre, Y., Breyton, C., Lemoine, Y., Robert B., Vernotte, C., Popot, J.L.: On the presence and role of a molecule of chlorophyll a in the cytochrome b 6 f complex. — J. Biol. Chem. 272: 21901–21908, 1997.

    Article  PubMed  CAS  Google Scholar 

  • Singh, A.: Interconversion of singlet oxygen and related species: Introduction. — Photochem. Photobiol. 28: 429–433, 1978.

    Article  Google Scholar 

  • Stroebel, D., Choquet, Y., Popot, J.L., Picot, D.: An atypical haem in the cytochrome b 6 f complex. — Nature 426: 413–418, 2003.

    Article  PubMed  CAS  Google Scholar 

  • Suh, H.J., Kim, C.S., Jung, J.: Cytochrome b 6 f complex as an indigenous photodynamic generator of singlet oxygen in thylakoid membranes. — Photochem. Photobiol. 71: 109–113, 2000.

    Article  Google Scholar 

  • Yan, J.S., Liu, Y.L., Mao, D.Z., Li, L.B., Kuang, T.Y.: The presence of 9-cis-β-carotene in cytochrome b6f complex from spinach. — Biochim. Biophys. Acta 1506: 182–188, 2001.

    Article  PubMed  CAS  Google Scholar 

  • Zhang, H., Joseph, J., Felix, C., Kalyanaraman B: Bicarbonate enhances the hydroxylation, nitration, and peroxidation reactions catalyzed by copper, zinc superoxide dismutase. Intermediacy of carbonate anion radical. — J. Biol. Chem. 275: 14038–14045, 2000.

    Article  PubMed  CAS  Google Scholar 

  • Zhang, H.M., Huang, D., Cramer, W.A.: Stoichiometrically bound β-carotene in the cytochrome b 6 f complex of oxygenic photosynthesis protects oxygen damage. — J. Biol. Chem. 274: 1581–1587, 1999.

    Article  PubMed  CAS  Google Scholar 

  • Zhang, S., Weng, J., Pan, J.X., Tu, T.C., Yao, S., Xu, C.H.: Study on the photogeneration of superoxide radicals in Photosystem II with EPR spin trapping techniques. — Photosynth. Res. 75: 41–48, 2003.

    Article  PubMed  CAS  Google Scholar 

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Acknowledgements

This work has been jointly supported by the National Basic Research Program of China (973 Programs, 2009CB118501 and 2009CB220008), the Knowledge Innovation Program of the Chinese Academy of Sciences (Grant No. KGCX2-YW-375-3 and KSCX2-YW-R-137), the National Natural Science Foundation (30900095) and the Natural Science Foundation of Hebei Province (C2008000684).

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Authors and Affiliations

  1. Institute of Hydrobiology, Ji’nan University, Guangzhou, 510632, P.R. China

    M. Sang, X. -C. Qin, W. -D. Wang, K. B. Wang, L. -B. Li & T. Y. Kuang

  2. Key Laboratory of Photosynthesis and Environmental Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing, 100093, P.R. China

    M. Sang

  3. Key Laboratory of Photochemistry, Institute of Chemistry, Chinese Academy of Sciences, Beijing, 100080, P.R. China

    J. Xie

  4. School of Bioscience and Engineering, Hebei University of Science and Technology, Shijiazhuang, Hebei Province, 050018, P.R. China

    X. -B. Chen

  5. Department of Chemistry, Renmin University of China, Beijing, 100872, P.R. China

    J. -P. Zhang

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  1. M. Sang
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  2. X. -C. Qin
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Additional information

An erratum to this article can be found at http://dx.doi.org/10.1007/s11099-011-0059-2

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Sang, M., Qin, X.C., Wang, W.D. et al. High-light-induced superoxide anion radical formation in cytochrome b 6 f complex from spinach as detected by EPR spectroscopy. Photosynthetica 49, 48–54 (2011). https://doi.org/10.1007/s11099-011-0008-0

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