Skip to main content
SpringerLink
Log in

The photoproduction of superoxide radicals and the superoxide dismutase activity of Photosystem II. The possible involvement of cytochrome b559

  • Regular paper
  • Published:
Photosynthesis Research Aims and scope Submit manuscript

Abstract

In the present study the light induced formation of superoxide and intrinsic superoxide dismutase (SOD) activity in PS II membrane fragments and D1/D2/Cytb559-complexes from spinach have been analyzed by the use of ferricytochrome c (cyt c(III)) reduction and xanthine/xanthine oxidase as assay systems. The following results were obtained: 1.) Photoreduction of Cyt c (III) by PS II membrane fragments is induced by addition of sodium azide, tetracyane ethylene (TCNE) or carbonylcyanide-p-trifluoromethoxy-phenylhydrazone (FCCP) and after removal of the extrinsic polypeptides by a 1M CaCl2-treatment. This activity which is absent in control samples becomes completely inhibited by the addition of exogenous SOD. 2.) The TCNE induced cyt c(III) photoreduction by PS II membrane fragments was found to be characterized by a half maximal concentration of c1/2=10 μM TCNE. Simultaneously, TCNE inhibits the oxygen evolution rate of PS II membrane fragments with c1/2≈ 3 μM. 3.) The photoproduction of O2 − is coupled with H+-uptake. This effect is diminished by the addition of the O2 −-trap cyt c(III). 4.) D1/D2/Cytb559-complexes and PS II membrane fragments deprived of the extrinsic proteins and manganese exhibit no SOD-activity but are capable of producing O2 − in the light if a PS II electron donor is added.

Based on these results the site(s) of light induced superoxide formation in PS II is (are) inferred to be located at the acceptor side. A part of the PS II donor side and Cyt b559 in its HP-form are proposed to provide an intrinsic superoxide dismutase (SOD) activity.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

ADRY:

acceleration of the deactivation reactions of the water-splitting system Y

ANT-2p:

2-(3-chloro-4-trifluoromethyl)anilino-3,5-dinitrothiophene

BCP:

bromocresol purple

cyt:

cytochrome

Cyt c:

cytochrome c

DCIP:

2,6-dichlorophenol-indophenol

DCMU:

3-(3,4-dichlorophenyl)-1,1-dimethylurea

DEDTC:

Diethyldithiocarbamate

DMBQ:

2,5-dimethyl-p-benzoquinone

DPC:

1,5-diphenylcarbazide

FCCP:

carbonylcyanide-p-trifluoro/methoxy-phenylhydrazone

HP:

high potential

LP:

low potential

MES:

2-(N-morpholino)ethanesulfonic acid

NADP:

nicotinamide adenine dinucleotide phosphate

SOD:

superoxide dismutase

TCNE:

tetracyane ethylene

TEMED:

N,N,N′,N′-tetramethylethylenediamine

References

  • Ananyev GM and Klimov VV (1988) Photoproduction of bound hydrogen peroxide in the donor side of Photosystem II. Dokl Akad Nauk SSSR 298: 1007–1011

    Google Scholar 

  • Ananyev GM and Klimov VV (1989) Study on the photoproduction of ‘bound’ hydrogen peroxide at the donor side of Photosystem II. Biokhimia USSR 54: 1587–1597

    Google Scholar 

  • Ananyev GM, Wydrzynski T, Renger G and Klimov VV (1992) Transient peroxide formation by the manganese-containing redox-active donor side of Photosystem II upon inhibition of O2 evolution with lauroylcholine chloride. Biochim Biophys Acta 1100: 303–311

    Google Scholar 

  • Armstrong DA and Buchanan JD (1978) Reaction of O2 −, H2O2 and other oxidants with sulfhydryl enzymes. Photochem Photobiol 28: 743–755

    PubMed  Google Scholar 

  • Arnon DJ and Barber J (1990) Photoreduction of NADP+ by isolated reaction centers of Photosystem II: Requirement for plastocyanin. Proc Natl Acad Sci USA 87: 5930–5934

    PubMed  Google Scholar 

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

    PubMed  Google Scholar 

  • Asada K and Takahashi M (1987) Production and scavenging of active oxygen in photosynthesis. In: Kyle DJ, Osmond CB and Arntzen CJ (eds) Photoinhibition, pp 227–287. Elsevier Science Publishers, Amsterdam

    Google Scholar 

  • Barber J, Chapman DJ and Telfer A (1987) Characterisation of a PS II reaction center isolated from the chloroplasts of Pisum sativum. FEBS Lett 220: 67–73

    Article  Google Scholar 

  • Bekina RM, Lebedeva AF und Shuvalov VA (1976) The reduction of oxygen in PS II with silicomolibdenic acid. Dokl Akad Nauk SSSR 231: 739–742

    Google Scholar 

  • Bendall DS (1982) Photosynthetic cytochromes of oxygenic organisms. Biochim Biophys Acta 683: 119–151

    Google Scholar 

  • Berthold DA, Babcock GT and Yocum CF (1981) A highly resolved, oxygen evolving Photosystem II preparation from spinach thylakoid membranes. FEBS Lett 134: 231–234

    Article  Google Scholar 

  • Bielski BHJ (1978) Reevaluation of the spectral and kinetic properties of HO2 and O2 − free radicals. Photochem Photobiol 28: 645–649

    Google Scholar 

  • Bolann BJ and Ulvik RJ (1993) Decay of superoxide catalysed by ferritin. FEBS Lett 318: 149–152

    Article  PubMed  Google Scholar 

  • Boucher N and Carpentier R (1993) Heat-stress stimulation of oxygen uptake by Photosystem I involves the reduction of superoxide radicals by specific electron donors. Photosynth Res 35: 213–218

    Article  Google Scholar 

  • Bowler C, Van Montagu M and Inze D (1992) Superoxide dismutase and stress tolerance. Annu Rev Plant Physiol Plant Mol Biol 43: 83–116

    Article  Google Scholar 

  • Bowyer JR and Camilleri P (1985) Spin-trap study of the reactions of ferrodoxin with reduced oxygen species in pea chloroplasts. Biochim Biophys Acta 808: 235–242

    Google Scholar 

  • Buser CA, Diner BA and Brudvig G (1992) Photooxidation of cytochrome b559 in oxygen evolving Photosystem II. Biochemistry 31: 11449–11459

    PubMed  Google Scholar 

  • Butler WL and Matsuda H (1983) Possible role of cytochrome b559 in Photosystem II. In: Inoue Y, Crofts AR, Govindjee, Murata N, Renger G and Satoh K (eds) The Oxygen Evolving System of Photosynthesis, pp 113–122. Academic Press, New York

    Google Scholar 

  • Canaani O and Havaux M (1990) Evidence for a biological role in photosynthesis for cytochrome b559 — a component of the Photosystem II reaction center. Proc Natl Acad Sci USA 87: 9295–9299

    PubMed  Google Scholar 

  • Chen G-X, Blubaugh DJ, Golback JH and Cheniae GM (1992) Photoinactivation of PS2 secondary donors by PS2 cation radicals and superoxide radicals. In: Murata N (ed) Research in Photosynthesis, Vol IV, pp 451–454. Kluwer Academic Publishers, Dordrecht

    Google Scholar 

  • Cramer WA and Whitmarsh J (1977) Photosynthetic cytochromes. Ann Rev Plant Physiol 28: 133–172

    Article  Google Scholar 

  • Cramer WA, Theg SM and Widger WR (1986) On the structure and function of cytochrome b559. Photosynth Res 10: 393–403

    Google Scholar 

  • Cross AR and Jones OTG (1991) Enzymatic mechanisms of superoxide production. Biochim Biophys Acta 1057: 281–298

    PubMed  Google Scholar 

  • Czapski G and Dorfman LM (1964) Pulse radiolysis studies. V. Transient spectra and rate constants in oxygenated aqueous solutions. J Phys Chem 68: 1169–1177

    Google Scholar 

  • Diner BA and Petrouleas V (1988) Q400 the non-heme iron of the Photosystem II iron-quinone complex. A spectroscopic probe of quinone and inhibitor binding to the reaction center. Biochim Biophys Acta 895: 107–125

    Google Scholar 

  • Eckert HJ, Wiese N, Bernarding J, Eichler HJ and Renger G (1988) Analysis of the electron transfer from Pheo− to QA in PS II membrane fragments from spinach by time resolved 325nm absorption changes in the picosecond time domain. FEBS Lett 240: 153–158

    Article  PubMed  Google Scholar 

  • Fischkoff S and Vanderkooi JM (1975) Oxygen Diffusion in Biological and Artificial Membranes Determined by the Fluorochrome Pyrene. J Gen Physiol 65: 663–676

    Article  PubMed  Google Scholar 

  • Ghanotakis DF, Babcock GT and Yocum CF (1984) Structural and catalytic properties of the oxygen evolving complex. Correlation of polypeptide and managanese release with the behaviour of Z+ in chloroplasts and a highly resolved preparation of the PS II complex. Biochim Biophys Acta 765: 388–398

    Google Scholar 

  • Ghanotakis DF, Yocum CF and Babcock GT (1986) ESR spectroscopy demonstrates that cytochrome b559 remains low potential in Ca2+-reactivated, salt-washed PS II particles. Photosynth Res 9: 125–134

    Google Scholar 

  • Goodman G and Leigh JS Jr. (1987) The distance between cytochromes a and a3 in the azide compound of bovine-heart cytochrome oxidase. Biochim Biophys Acta 890: 360–367

    PubMed  Google Scholar 

  • Hewitt EJ and Nicholas DJD (1963) Cations and Anions: Inhibition and Interactions in Metabolism and in Enzyme Activity. In: Metabolic Inhibitors, Vol II, Ch. 29, Hochster RM and Quastel JH (eds.). Academic Press, New York and London

    Google Scholar 

  • Hideg E, Kobayashi M and Inaba H (1991) Spontaneous ultraweak light emission from respiring spinach leaf mitochondria. Biochim Biophys Acta 1098: 27–31

    Google Scholar 

  • Ilan YA, Czapski G and Meisel D (1976) The one-electron transfer redox potentials of free radicals. I. The oxygen/superoxide system. Biochim Biophys Acta 430: 209–224

    PubMed  Google Scholar 

  • Irrgang K-D (1990) Untersuchungen zur Polypeptidstruktur des Photosystems II aus Höheren Pflazen. Dissertation TU-Berlin, D83

  • Irrgang K-D, Geiken B, Lange B and Renger G (1992) Disulfide bridge modifiers and sulfhydryl group blockers are inactivating the oxygen evolving enzyme of PS II from spinach. In Murata N (ed) Research in Photosynthesis, Vol II, pp 417–420. Kluwer Academic Publishers. Dordrecht

    Google Scholar 

  • Jursinic PA (1978) Flash polarographic detection of superoxide production as a means of monitoring electron flow between Photosystem I and II. FEBS Lett 90: 15–20

    Article  Google Scholar 

  • Kanematsu S and Asada K (1989) CuZn-superoxide dismutases in rice: Occurence of an active, monomeric enzyme and two types of isozyme in leaf and non-photosynthetic tissues. Plant Cell Physiol. 30: 381–391

    Google Scholar 

  • Klimov VV, Allakhverdiev SJ, Demeter S and Krasnovsky AA (1979) Photoreduction of pheophytin in Photosystem II of chloroplasts as a function of the redox potential of the medium. Dokl Akad Nauk SSSR 249: 227–230

    Google Scholar 

  • Klimov VV, Ananyev GM, Zastryzhnaya OM, Wydryzynski T and Renger G 1993 Photoproduction of hydrogen peroxide in Photosystem II membrane fragments: A comparison of four signals Photosynth Res 38: 409–416

    Google Scholar 

  • Koppenol WH, Van Buuren KJH, Butler J and Braams R (1976) The kinetics of the reduction of cytochrome c by the superoxide radical. Biochim Biophys Acta 449: 157–168

    PubMed  Google Scholar 

  • Koulogliotis D, Kostopoulos T, Petrouleas V and Diner B (1993) Evidence for CN− binding at the PS II non-heme Fe2+. Effects on the EPR signal for QA Fe2+ and on QA/QB electron transfer. Biochim Biophys Acta 1141: 275–282

    Google Scholar 

  • Matsuda H and Butler WL (1983) Restoration of high potential cytochrome b559 in Photosystem II particles and liposomes. Biochim Biophys Acta 725: 320–324

    Google Scholar 

  • Mc Cord JM and Fridovich I (1969) Superoxide dismutase. An enzymic function for erythrocuprein (hemocuprein). J Biol Chem 244: 6049–6055

    PubMed  Google Scholar 

  • Mehler AH (1951) Studies on reactions of illuminated chloroplasts. I. Mechanism of the reduction of oxygen and other Hill reagents. Arch Biochem Biophys 33: 65–77

    Google Scholar 

  • Mei R and Yocum CF (1992) Comparative properties of hydroquinone and hydroxylamine reduction of the Ca2+ stabilized O2-evolving complex of Photosystem II: Reductant-dependent Mn2+ formation and activity inhibition. Biochemistry 31: 8449–8454

    PubMed  Google Scholar 

  • Misra HP (1979) Reaction of copper-zinc superoxide dismutase with diethyldithiocarbamate. J Biol Chem 254: 11623–11628

    PubMed  Google Scholar 

  • Mizusawa N and Yamashita T (1992) The shift of redox potentials Eo′ of cytochrome b559 through the photo-reactivation of the oxygen evolving center. In: Murata N (ed) Research in Photosynthesis, Vol II, pp 425–428. Kluwer Academic Publishers, Dordrecht

    Google Scholar 

  • Nanba O and Satoh K (1987) Isolation of a Photosystem II reaction center consisting of D1 and D2 polypeptides and cytochrome b559. Proc Natl Acad Sci USA 84: 109–112

    Google Scholar 

  • Nuijs AM, van Gorkom HJ, Plijter JJ and Duysens LMN (1986) Primary charge separation and excitation of chlorophyll a in Photosystem II particles from spinach as studied by picosecond absorbance-difference spectroscopy. Biochim Biophys Acta 848: 167–175

    Google Scholar 

  • Oh-oka H, Tanaka S, Wada K, Kuwabara T and Murata N (1986) Complete amino acid sequence of the 33 kDa protein isolated from spinach Photosystem II particles. FEBS Lett 197: 63–66

    Article  Google Scholar 

  • Ortega JM, Hervas M and Losada M (1992) A possible role for cytochrome b559 in photosynthesis. In: Murata N (ed) Research in Photosynthesis, Vol II, pp 697–700. Kluwer Academic Publishers, Dordrecht

    Google Scholar 

  • Rabinowitch HD and Fridovich I (1983) Superoxide radicals, superoxide dismutases and oxygen toxicity in plants. Photochem Photobiol 37: 679–690

    Google Scholar 

  • Renger G (1972) The action of 2-anilinothiophenes as accelerators of the deactivation reactions in the water-splitting enzyme system of photosynthesis. Biochim Biophys Acta 256: 428–439

    PubMed  Google Scholar 

  • Renger G (1987) Biological Exploitation of Solar Energy by Photosynthetic water splitting. Angew Chem Int Ed Engl 26: 643–660

    Article  Google Scholar 

  • Renger G, Bouges-Bocquet B and Delosme R (1973) Studies on the ADRY agent induced mechanism of the discharge of the holes trapped in the photosynthetic watersplitting enzyme system Y. Biochim Biophys Acta 292: 796–807

    PubMed  Google Scholar 

  • Renger G, Wacker U and Völker M (1987) Studies on the protolytic reactions coupled with water cleavage in Photosystem II membrane fragments from spinach. Photosynth Res 13: 167–189

    Google Scholar 

  • Samson G and Fork DC (1992) Simultaneous photoreduction and photooxidation of cytochrome b559 in Photosystem II treated with carbonylcyanide-m-chlorophenylhydrazone. Photosynth Res 33: 203–212

    Google Scholar 

  • Sayre RT and Homann PH (1979) A light dependent oxygen consumption induced by Photosystem II of isolated chloroplasts. Arch Biochem Biophys 196: 525–533

    PubMed  Google Scholar 

  • Schröder WP and Åkerlund H-E (1986) H2O2 accessibility to the Photosystem II donor side in protein-depleted inside out thylakoids measured as flash-induced oxygen production. Biochim Biophys Acta 848: 359–363

    Google Scholar 

  • Schröder WP and Åkerlund H-E (1990) Hydrogen peroxide production in Photosystem II preparations. In: Baltscheffsky M (ed) Current Research in Photosynthesis, Vol I, pp 901–904. Kluwer Academic Publishers, Dordrecht

    Google Scholar 

  • Seibert M, De Wit M and Staehelin LA (1987) Structural localization of the O2-evolving apparatus to multimeric (tetrameric) particles on the lumenal surface of freeze-etched photosynthetic membranes. J. Cell Biol. 105: 2257–2265

    Article  PubMed  Google Scholar 

  • Seki H, Ilan YA, Stein G (1976) Reactions of the ferriferrocytochrome c system with superoxide/oxygen and CO2 −/CO2 studied by fast pulse radiolyses. Biochim Biophys Acta 440: 573–586

    PubMed  Google Scholar 

  • Shiraishi T, Takahashi M and Asada K (1992) Production and decay of superoxide radicals in illuminated thylakoid membranes. In: Murata N (ed) Research in Photosynthesis, Vol II pp 627–634. Kluwer Academic Publishers, Dordrecht

    Google Scholar 

  • Shuvalov VA and Krasnovsky AA (1975) Investigation of the photoreduction of oxygen in chloroplasts measured as chemiluminescence of luminol and chlorophyll. Biokhimia USSR 40: 358–367

    Google Scholar 

  • Shuvalov VA, Heber U and Schreiber U (1989) Low temperature photochemistry and spectral properties of a Photosystem II reaction center complex containing the proteins D1 and D2 and two hemes of cytochrome b559. FEBS Lett 258: 27–31

    Article  Google Scholar 

  • Takahashi M and Asada K (1988) Superoxide production in the aprotic interior of chloroplast thylakoids. Arch Biochem Biophys 267: 714–722

    PubMed  Google Scholar 

  • Tanaka S and Wada K (1988) The status of cysteine residues in the extrinsic 33 kDa protein of spinach Photosystem II complexes. Photosynth Res 17: 255–266

    Google Scholar 

  • Thompson LK and Brudvig GW (1988) Cytochrome b559 may function to protect Photosystem II from photoinhition. Biochemistry 27: 6653–6658

    PubMed  Google Scholar 

  • Van Gelder (1962) The extinction coefficient of cytochrome c. Biochim Biophys Acta 58: 593–595

    Article  PubMed  Google Scholar 

  • Völker M, Renger G and Rutherford M (1986) Effects of trypsin upon EPR-signals arising from components of the donor side of Photosystem II. Biochem Biophys Acta 851: 424–430

    Google Scholar 

  • Wacker U, Haag E and Renger G (1990) Investigation of pH-change patterns of Photosystem II membrane fragments from spinach. In: Baltscheffsky M (ed) Current Research in Photosynthesis, Vol I, pp 869–872. Kluwer Academic Publishers, Dordrecht

    Google Scholar 

  • Wacker U (1993) Untersuchung der Kopplung von H+-Reaktionen und Elektronentransfervorgängen im Photosystem II Höherer Pflanzen. Dissertation TU-Berlin, D83

  • Wood M (1974) The redox potential of the system oxygen-superoxide. FEBS Lett 44: 23–24

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institut of Soil Science and Photosynthesis, Russian Academy of Sciences, Pushchino, Moskow Region, (Russia)

    Gennady Ananyev & Vyacheslav Klimov

  2. Max-Volmer Institut für Biophysikalische und Physikalische Chemie Technische Universität Berlin, 10623, Berlin, (Germany)

    Gernot Renger & Ulrich Wacker

Authors
  1. Gennady Ananyev
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Gernot Renger
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ulrich Wacker
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Vyacheslav Klimov
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ananyev, G., Renger, G., Wacker, U. et al. The photoproduction of superoxide radicals and the superoxide dismutase activity of Photosystem II. The possible involvement of cytochrome b559. Photosynth Res 41, 327–338 (1994). https://doi.org/10.1007/BF00019410

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00019410

Key words

Search

Navigation