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MOA-stilbene: A new tool for investigation of the reactions of the chloroplast cytochrome bf complex

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Abstract

MOA-stilbene is known to be a specific inhibitor of the Qo site of mammalian cytochrome bc 1 complex. We show that it also binds to the chloroplast cytochrome bf complex. Binding to the reduced enzyme induces a red-shift of the Soret and visible absorption bands of the haems b. Steady state and single turnover experiments with thylakoid membranes show that MOA-stilbene promotes additional ‘oxidant-induced reduction’ of the b haems and slows their subsequent dark reoxidation. In single turnover experiments, the associated slow phase of the carotenoid bandshift at 518 nm is only partially decreased in apparent extent and rate. These and other effects are similar to those produced by NQNO, a Qi site effector, and by analogy indicate that MOA-stilbene should also be primarily a Qi-site effector of the cytochrome bf complex. MOA-stilbene has less effect on other parts of the photosynthetic chain. This confers an important advantage on MOA-stilbene in that its effects on the cytochrome bf complex can be studied by using Photosystem II to activate turnover. Myxothiazol displays effects on the cytochrome bf complex which are similar to, but much weaker than, those of MOA-stilbene.

A Q cycle-based model of turnover of the cytochrome bf complex is presented, which can account for several unusual features of kinetic behaviour.

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Abbreviations

DBMIB:

2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone

duroquinol:

2,3,5,6-tetramethyl-p-benzohydroquinone

Ehx :

Ambient potential at pHx versus SHE

Emx :

Midpoint potential at pH x versus SHE

haem b H :

the higher potential haem b of cytochrome b, thought to be associated with the quinone reduction site, Qi, and sometimes termed haem b n

haem b L :

the lower potential haem of cytochrome b, thought to be associated with the quinol oxidation site, Qo, and sometimes termed haem b p

HQNO:

2-n-heptyl-4-hydroxyquinoline-N-oxide

MOA-stilbene:

E-β-methoxyacrylate-stilbene or (E,E)-methyl 3-methoxy-2-(styrylphenyl)propenoate

NQNO:

2-n-nonyl-4-hydroxyquinoline-N-oxide

QB (site):

the (binding site of the) secondary quinone acceptor of Photosystem II

Qo site:

the quinol oxidation site and site of proton output of the bc and bf complexes (also termed the Qz or Qp site)

Qi site:

the quinone reduction site and site of proton input of the bc and bf complexes (also termed the Qc, Qr or Qn site)

SHE:

Standard Hydrogen Electrode

References

  • Avron M (1961) The inhibition of photoreactions in chloroplasts by 2-alkyl-4-hydroxyquinoline N-oxides. Biochem J 78: 735–739

    Google Scholar 

  • Beautement K, Clough JM, de Fraine PJ and Godfrey CRA (1991) Fungicidal β-methoxyacrylates: From natural products to novel synthetic agricultural fungicides. Pesticide Science 31: 499–519

    Google Scholar 

  • Brandt U, Schägger H and von Jagow G (1988) Characterisation of binding of the methoxyacrylate inhibitors to mitochondrial cytochrome c reductase. Eur J Biochem 173: 499–506

    Google Scholar 

  • Clark WM (1960) Oxidation-Reduction Potentials of Organic Systems. Bailliere, Tindall & Cox, Ltd., London

    Google Scholar 

  • Cramer WA, Black MT, Widger WR and Girvin ME (1987) Structure and function of photosynthetic b-c 1 and b 6-f complexes. In: Barber J (ed) The Light Reactions, pp 447–493. Elsevier Science Publishers, Amsterdam

    Google Scholar 

  • Delosme R, Joliot P and Trebst A (1987) Flash-induced oxidation of cytochrome b-563 in algae under anaerobic conditions: Effect of dinitrophenylether of iodonitrothymol. Biochim Biophys Acta 893: 1–6

    Google Scholar 

  • Dutton PL and Wilson DF (1974) Redox potentiometry in mitochondrial and photosynthetic bioenergetics. Biochim Biophys Acta 346: 165–212

    Google Scholar 

  • Hartung A and Trebst A (1985) New inhibitiors of the tochrome b 6/f-complex in ferredoxin catalyzed cyclic photophosphorylation. Physiologie Vegetale 23: 635–648

    Google Scholar 

  • Hauska G, Nitschke W and Herrmann RG (1988) Amino acid identities in the three redox center-carrying polypeptides of cytochrome bc 1/b 6 f complexes. J Bioenerg Biomemb 20: 211–228

    Google Scholar 

  • Hope AB and Matthews DB (1987) The slow phase of the electrochromic shift in relation to the Q-cycle in chloroplasts. Aust J Plant Physiol 14: 29–46

    Google Scholar 

  • Hope AB and Rich PR (1989) Proton uptake by the chloroplast cytochrome bf complex. Biochim Biophys Acta 975: 96–103

    Google Scholar 

  • Hope AB, Liggins J and Matthews DB (1989) The kinetics of reactions in and near the cytochrome b/f complex of chloroplasts. II Cytochrome b-563 reduction. Aust Journ Plant Physiol 16: 353–364

    Google Scholar 

  • Hurt E and Hauska G (1981) A cytochrome f/b 6 complex of five polypetides with plastoquinol-plastocyanin-oxidoreductase activity from spinach chloroplasts. Eur J Biochem 117: 591–599

    Google Scholar 

  • Joliot P and Joliot A (1984) Electron transfer between the two Photosystems I. Flash excitation under oxidizing conditions. Biochim Biophys Acta 765: 210–218

    Google Scholar 

  • Joliot P, Lavergne J and Béal D (1990) Organisation of the plastoquinone pool in chloroplasts: Evidence for clusters of different sizes. In: Baltscheffsky M (ed) Current Research in Photosynthesis, pp 879–882. Kluwer Academic Publishers, Dordrecht

    Google Scholar 

  • Jones RW and Whitmarsh J (1985) Origin of the electrogenic reaction in the chloroplast cytochrome b/f complex. Photobiochem Photobiophys 9: 119–127

    Google Scholar 

  • Jones RW and Whitmarsh J (1988) Inhibition of electron transfer and the electrogenic reaction in the cytochrome b/f complex by 2-n-nonyl-4-hydroxyquinoline N-oxide (NQNO) and 2,5-dibromo-3-methyl-6-isopropyl-p-benquinone (DBMIB). Biochim Biophys Acta 933: 258–268

    Google Scholar 

  • Lam E (1984) The effects of quinone analogues on cytochrome b 6 reduction and oxidation in a reconstituted system. FEBS Lett 172: 255–260

    Google Scholar 

  • Lam E and Malkin R (1982a) Ferredoxin-mediated reduction of cytochrome b-563 in a chloroplast cytochrome b-563/f complex. FEBS Lett 141: 98–101

    Google Scholar 

  • Lam E and Malkin R (1982b) In vitro reconstitution of the electron pathway from water to cytochrome f of spinach thylakoids. FEBS Lett 144: 190–193

    Google Scholar 

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

    Google Scholar 

  • Moss DA and Bendall DS (1984) Cyclic electron transport in chloroplasts. The Q-cycle and the site of action of antimycin. Biochim Biophys Acta 767: 389–395

    Google Scholar 

  • O'Keefe DP (1983) Sites of cytochrome b-563 reduction, and the mode of action of DNP-INT and DBMIB in the chloroplast cytochrome b-563/f complex. FEBS Lett 162: 349–354

    Google Scholar 

  • Oettmeier W, Godde D, Kunze B and Höfle G (1985) Stigmatellin. A dual type inhibitor of photosynthetic electron transport. Biochim Biophys Acta 807: 216–219

    Google Scholar 

  • Pace RJ, Hope AB and Smith P (1992) Detection of flashinduced quinone radicals in spinach chloroplasts. Biochim Biophys Acta 1098: 209–216

    Google Scholar 

  • Rich PR (1984) Electron and proton transfers through quinones and cytochrome bc complexes. Biochim Biophys Acta 768: 53–79

    Google Scholar 

  • Rich PR (1988) A critical examination of the supposed variable proton stoichiometry of the chloroplast cytochrome bf complex. Biochim Biophys Acta 932: 33–42

    Google Scholar 

  • Rich PR and Wiggins TE (1992) Soret and visible band circular dichroism measurements on the cytochrome bc 1 and bf complexes. Biochem Soc Trans 20: 2415

    Google Scholar 

  • Rich PR, Heathcote P and Moss DA (1987) Kinetic studies of electron transfer in a hybrid system constructed from the cytochrome bf complex and Photosystem I. Biochim Biophys Acta 892: 138–151

    Google Scholar 

  • Rich PR, Jeal AE, Madgwick SA and Moody AJ (1990) The effects of inhibitors on the redox linked protonations of the haems b of the mitochondrial bc 1 complex. Biochim Biophys Acta 1018: 29–40

    Google Scholar 

  • Rich PR, Madgwick SA and Moss DA (1991) The interactions of duroquinol, DBMIB and NQNO with the chloroplast cytochrome bf complex. Biochim Biophys Acta 1058: 312–328

    Google Scholar 

  • Selak MA and Whitmarsh J (1982) Kinetics of the electrogenic step and cytochrome b 6 and f redox changes in chloroplasts. Evidence for a Q cycle. FEBS Lett 150: 286–292

    Google Scholar 

  • Trebst A (1980) Inhibitors of electron flow: Tools for the functional and structural localization of carriers and energy conservation sites. In: Colowick SP and Kaplan NO (eds) Methods in Enzymology, Vol 69c, pp 675–715. Academic Press, New York

    Google Scholar 

  • Trebst A and Wietoska H (1989) On an inhibitor of ferredoxin catalyzed cyclic photophosphorylation in thylakoid membranes. Botanica Acta 102: 302–305

    Google Scholar 

  • Trebst A, Wietoska H, Draber W and Knops HJ (1978) The inhibition of photosynthetic electron flow in chloroplasts by the dinitrophenylether of bromo-or iodo-nitrothymol. Z Naturforsch 33: 919–927

    Google Scholar 

  • von Jagow G and Link TA (1986) Use of specific inhibitors on the mitochondrial bc 1 complex. Meth Enzymol 126: 253–271

    Google Scholar 

  • von Jagow G and Ohnishi T (1985) The chromone inhibitor stigmatellin-binding to the ubiquinol oxidation center at the C-side of the mitochondrial membrane. FEBS Lett 185: 311–315

    Google Scholar 

  • Witt HT (1979) Energy conversion in the functional membrane of photosynthesis. Analysis by light pulse and electric field methods. The central role of the eletric field. Biochim Biophys Acta 505: 355–427

    Google Scholar 

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Authors and Affiliations

  1. Glynn Research Institute, PL30 4AU, Bodmin, Cornwall, United Kingdom

    Peter R. Rich, Sally A. Madgwick & Simon Brown

  2. Gustav-Embden-Zentrum der Biologischen Chemie, Klinikum der Johann Wolfgang Goethe-Universität, 6000, Frankfurt, Germany

    Gebhard von Jagow & Ulrich Brandt

Authors
  1. Peter R. Rich
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  2. Sally A. Madgwick
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  3. Simon Brown
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  4. Gebhard von Jagow
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  5. Ulrich Brandt
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Rich, P.R., Madgwick, S.A., Brown, S. et al. MOA-stilbene: A new tool for investigation of the reactions of the chloroplast cytochrome bf complex. Photosynth Res 34, 465–477 (1992). https://doi.org/10.1007/BF00029819

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  • DOI: https://doi.org/10.1007/BF00029819

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