Skip to main content
SpringerLink
Log in

Photoinhibition of carotenoidless reaction centers from Rhodobacter sphaeroides by visible light. Effects on protein structure and electron transport

  • Published:
Photosynthesis Research Aims and scope Submit manuscript

Abstract

Inhibition of electron transport and damage to the protein subunits by visible light has been studied in isolated reaction centers of the non-sulfur purple bacterium Rhodobacter sphaeroides. Illumination by 1100 μEm−2 s−1 light induced only a slight effect in wild type, carotenoid containing 2.4.1. reaction centers. In contrast, illumination of reaction centers isolated from the carotenoidless R26 strain resulted in the inhibition of charge separation as detected by the loss of the initial amplitude of absorbance change at 430 nm arising from the P+QB − → PQB recombination. In addition to this effect, the L, M and H protein subunits of the R26 reaction center were damaged as shown by their loss on Coomassie stained gels, which was however not accompanied by specific degradation products. Both the loss of photochemical activity and of protein subunits were suppressed in the absence of oxygen. By applying EPR spin trapping with 2,2,6,6-tetramethylpiperidine we could detect light-induced generation of singlet oxygen in the R26, but not in the 2.4.1. reaction centers. Moreover, artificial generation of singlet oxygen, also led to the loss of the L, M and H subunits. Our results provide evidence for the common hypothesis that strong illumination by visible light damages the carotenoidless reaction center via formation of singlet oxygen. This mechanism most likely proceeds through the interaction of the triplet state of reaction center chlorophyll with the ground state triplet oxygen in a similar way as occurs in Photosystem II.

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

References

  • Andersson B and Styring S (1991) Photosystem II: Molecular organization, function and acclimation. Curr Top Bioenerg 16: 1–81

    CAS  Google Scholar 

  • Andersson B, Salter H, Virgin I, Vass I and Styring S (1992) Photodamage to Photosystem II - primary and secondary events.J Photochem Photobiol 15 B: 15–31

    Google Scholar 

  • Aro E-M, Virgin I and Andersson B (1993) Photoinhibition of Photosystem II. Inactivation, protein damage and turnover. Biochim Biophys Acta 1143: 113–134

    Article  PubMed  CAS  Google Scholar 

  • Barber J (1998) Photosystem two. Biochim Biophys Acta 1365: 269–277

    Article  PubMed  CAS  Google Scholar 

  • Boucher F, Rest MVD and Gingras G (1977) Structure and function of carotenoids in the photoreaction center from Rhodospirillum rubrum. Biochim Biophys Acta 461: 339–357

    Article  PubMed  CAS  Google Scholar 

  • Brzezinski P and Andreasson LE (1995) Trypsin treatment of reaction centers from Rhodobacter sphaeroides in the dark and under illumination: Protein structural changes follow charge separation. Biochemistry 34: 7498–7506

    Article  PubMed  CAS  Google Scholar 

  • Chang C-H, Tiede D, Tang J, Smith U, Norris J and Schiffer M (1986) Structure of Rhodopseudomonas sphaeroides R-26 reaction center. FEBS Lett 205: 82–86

    Article  PubMed  CAS  Google Scholar 

  • Cleland RE, Rees D and Horton P (1992) Light-induced fluorescence quenching and loss of photochemistry in chromatophores of photosynthetic purple bacteria. J Photochem Photobiol 13: 253–265

    Article  CAS  Google Scholar 

  • Cogdell RJ and Frank HA (1987) How carotenoids function in photosynthetic bacteria. Biochim Biophys Acta 895: 63–79

    PubMed  CAS  Google Scholar 

  • di Mascio P, Kaiser S and Sies H (1989) Lycopene as the most efficient biological carotenoid singlet oxygen quencher. Arch Biochem Biophys 274: 532–538

    Article  PubMed  CAS  Google Scholar 

  • Durrant JR, Giorgi LB, Barber J, Klug DR, and Porter G (1990) Characterization of triplet states in isolated Photossytem II reaction centres: Oxygen quenching as a mechanism for photodamage. Biochim Biophys Acta 1017: 167–175

    Article  CAS  Google Scholar 

  • Eijckelhoff C and Dekker JP (1995) Determination of the pigment stoichiometry of the photochemical reaction center of Photosystem II. Biochim Biophys Acta 1231: 21–28

    Article  Google Scholar 

  • Ermler U, Fritzsch G, Buchanan SK and Michel H (1994) Structure of the photosynthetic reaction centre from Rb. sphaeroides at 2.65 Å resolution: Cofactors and protein-cofactor interactions. Structure 2: 925–936

    Article  PubMed  CAS  Google Scholar 

  • Farhoosh R, Chynwat V, Gebhard R, Lugtenburg J and Frank HA (1997) Triplet energy transfer between the primary donor and carotenoids in Rhodobacter sphaeroides R-26.1 reaction centers incorporated with Spheroidene analogs having different extents of pi-electron conjugation. Photochem Photobiol 66: 97–104

    PubMed  CAS  Google Scholar 

  • Foote CS (1968) Photosensitized oxygenations and the role of singlet O2. Acc Chem Res 1: 104–110

    Article  CAS  Google Scholar 

  • Frank HA and Cogdell RJ (1996) Carotenoids in photosynthesis. Photochem Photobiol 63: 257–264

    PubMed  CAS  Google Scholar 

  • Frank HA and Violette CA (1989) Monomeric bateriochlorophyll is required for the triplet energy transfer between the primary donor and the carotenoid in photosynthetic bacterial reaction centers. Biochim Biophys Acta 976: 222–232

    PubMed  CAS  Google Scholar 

  • Frank HA, Machnicki J and Friesner R (1983) Energy transfer between the primary donor bacteriochlorophyll and carotenoids in Rhodopseudomonas sphaeroides. Photochem Photobiol 38: 451–455

    CAS  Google Scholar 

  • Frank HA, Chynwat V, Hartwich G, Meyer M, Katheder I and Scheer H (1993) Carotenoid triplet state formation in Rhodobacter sphaeroides R-26 reaction centers exchanged with modified bacteriochlorophyll pigments and reconstituted with spheroidence. Photosynth Res 37: 193–203

    Article  CAS  Google Scholar 

  • Griffiths M, Sistrom WR, Cohen-Bazire G, and Stanier RY (1995) Functions of carotenoids in photosynthesis. Nature 176: 1211–1214

    Article  Google Scholar 

  • Hanley J, Deligiannakis Y, Pascal A, Faller P and Rutherford AW (1999) Carotenoid oxidation in Photosystem II. Biochemistry 38: 8189–8195

    Article  PubMed  CAS  Google Scholar 

  • Hideg É, Spetea C and Vass I (1994) Singlet oxygen and free radical poduction during acceptor-and donor-side-induced photoinhibition. Studies with spin trapping EPR spectroscopy. Biochim Biophys Acta 1186: 143–152

    Article  CAS  Google Scholar 

  • Hideg É, Spetea C and Vass I (1994) Singlet oxygen production in thylakoid membranes during photoinhibition as detected by EPR spectroscopy. Photosynth Res 39: 191–199

    Article  CAS  Google Scholar 

  • Kálman L, LoBrutto R, Allen JP and Williams JC (1999) Modified reaction centres oxidize tyrosine in reactions that mirror PS II. Nature 402: 696–699

    Article  Google Scholar 

  • Kleinfeld D, Okamura MY and Feher G (1984) Electron transfer in reaction centers of Rhodopseumonas sphaeroides. I. Determination of the charge recombination pathway of D+QAQ— B and free energy and kinetic relations between Q— A QB and QAQ— B. Biochim Biophys Acta 766: 126–140

    Article  PubMed  CAS  Google Scholar 

  • Krinsky NI (1968) The protective function of carotenoid pigments. In: Giese AC (ed) Photophysiology, pp 123–195. Academic Press, New York

    Google Scholar 

  • Kyle DJ, Ohad I and Arntzen CJ (1984) Membrane protein damage and repair: Selective loss of a quinone-protein function in chloroplast membranes. Proc Natl Acad Sci USA 81: 4070–4074

    Article  PubMed  CAS  Google Scholar 

  • Macpherson AN, Telfer A, Barber J and Truscott TG (1993) Direct detection of singlet oxygen from Photosystem II reaction centres. Biochim Biophys Acta 1143: 301–309

    Article  CAS  Google Scholar 

  • Maróti P and Wraight CA (1988) Flash-induced H+ binding by bacterial photosynthetic reaction centers: Comparison of spectrophotometric and conductimetric methods. Biochim Biophys Acta 934: 314–328

    Article  Google Scholar 

  • Mathis P (1994) Electron transfer between cytochrome c2 and the isolated reaction center of the purple bacterium Rhodobacter sphaeroides. Biochim Biophys Acta 1187: 177–180

    Article  CAS  Google Scholar 

  • Michel H and Deisenhofer J (1988) Relevance of the photosynthetic reaction center from purple bacteria to the structure of Photosystem II. Biochemistry 27: 1–7

    Article  CAS  Google Scholar 

  • Nagy L, Puskas A, Tandori J, Droppa M and Horvath G (1991) Effect of DCMU on photosynthetic purple bacteria. Photosynthetica 25: 167–171

    CAS  Google Scholar 

  • Ohad I, Kyle DJ and Arntzen CJ (1984) Membrane protein damage and repair: Removal and replacement of inactivated 32-kilodalton polypeptides in chloroplast membranes. J Cell Biol 99: 481–485

    Article  PubMed  CAS  Google Scholar 

  • Prasil O, Adir N and Ohad I (1992) Dynamics of Photosystem II: mechanism of photoinhibition and recovery processes. In: Barber J (ed) Topics in Photosynthesis, Vol 11: The Photosystems: Structure, Function and Molecular Biology, pp 295–348. Elsevier, Amsterdam

    Google Scholar 

  • Rosen D, Okamura MY and Feher G (1980) Interaction of Cytochrome c with reaction centers of Rhodopseudomonas sphaeroides R-26: Determination of number of binding sites and dissociation constants by equilibrium dialysis. Biochemistry 19: 5687–5692

    Article  PubMed  CAS  Google Scholar 

  • Rutherford AW (1986) How close is the analogy between the reaction centre of Photosystem II and that of purple bacteria ? Biochem Soc Trans 14: 15–17

    PubMed  CAS  Google Scholar 

  • Schenck CC, Mathis P and Lutz M (1984) Triplet formation and triplet decay in reaction centers from the photosynthetic bacterium Rhodopseudomonas spharoides. Photochem Photobiol 39: 407–417

    CAS  Google Scholar 

  • Shinkarev VP and Wraight CA (1993) Kinetic factors in the bicycle model of oxygen evolution by Photosystem II. Photosynth Res 38: 315–321

    Article  CAS  Google Scholar 

  • Takiff L and Boxer SG (1988) Phosphorescence spectra of bacteriochlorophylls. J Am Chem Soc 110: 4425–4426

    Article  CAS  Google Scholar 

  • Telfer A, Rivas DL and Barber J (1991) β-Carotene within the isolated Photosystem II reaction centre: Photooxidation and irreversible bleaching of this chromophore by oxidised P680. Biochim Biophys Acta 1060: 106–114

    Article  CAS  Google Scholar 

  • Vass I, Styring S, Hundal T, Koivuniemi A, Aro E-M and Andersson B (1992) Reversible and irreversible intermediates during photoinhibition of Photosystem II: Stable reduced QA species promote chlorophyll triplet formation. Proc Natl Acad Sci USA 89: 1408–1412

    Article  PubMed  CAS  Google Scholar 

  • Wraight CA and Stein RR (1983) Bacterial reaction centers as a model for Photosystem II: Turnover of the secondary acceptor quinone. In: Inoue Y, Crofts AR, Govindjee, Murata N, Renger G and Satoh K (eds) The Oxygen Evolving System of Photosynthesis, pp 383–391. Academic Press, Tokyo

    Google Scholar 

  • Yeates TO, Komiya H, Chirino A, Rees DC, Allen JP and Feher G (1988) Structure of the reaction center from Rhodobacter sphaeroides R-26 and 2.4.1: Protein-cofactor (bacteriochlorophyll, bacteriopheophytin and carotenoid) interactions. Proc Natl Acad Sci USA 85: 7993–7997

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Biophysics, University of Szeged, Hungary

    Júlia Tandori, László Nagy & Péter Maróti

  2. Institute of Plant Biology, Biological Research Center, P.O. Box 521, H-6701, Szeged, Hungary

    Éva Hideg & Imre Vass

Authors
  1. Júlia Tandori
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Éva Hideg
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. László Nagy
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Péter Maróti
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Imre Vass
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Imre Vass.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Tandori, J., Hideg, É., Nagy, L. et al. Photoinhibition of carotenoidless reaction centers from Rhodobacter sphaeroides by visible light. Effects on protein structure and electron transport. Photosynthesis Research 70, 175–184 (2001). https://doi.org/10.1023/A:1017907404325

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1017907404325

Search

Navigation