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Localization of light-induced structural changes in bacterial photosynthetic reaction centers

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Abstract

Light-induced structural changes in photosynthetic reaction centers from Rhodobacter sphaeroides were investigated using two approaches. Cu2+ was used as a paramagnetic structural probe. The EPR spectrum of Cu2+ incorporated into the metal-depleted reaction centers was affected by 1,10-phenanthroline, an electron transfer inhibitor substituting QB, which suggests a localization of Cu2+ in a vicinity of the Q −B site. However, the spectrum was not influenced by low temperature (77 K) illumination of the sample which suggests that the copper ion position is not exactly the same as that of the iron ion. Freezing the reaction centers under illumination in the presence of potassium ferricyanide and 1,10-phenanthroline caused a change in the shape of the Cu2+ EPR spectrum in comparison to that of a sample frozen in darkness. These data indicate a change of the Cu2+ ligand symmetry owing to light-induced structural changes which are probably located near the acceptor side of the reaction center. Partial trypsinolysis of reaction centers was also used to locate the structural changes. Trypsin treatment in the dark and under illumination resulted in different peptide patterns as detected by gel electrophoresis and reverse-phase high-performance liquid chromatography. Partial amino-acid sequence analysis of a number of peptides, characteristic of either light- or dark-treated reaction centers, showed that they originated from the acceptor sides of the H and M subunits. The occurrence of light-induced structural differences in the H-subunit is consistent with the suggestion that it may be involved in regulating electron transfer in this part of the reaction center.

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References

  • Allen JP, Feher G, Yeates TO, Komiya H and Rees DC (1988) Structure of the reaction center from Rhodobacter sphaeroides R-26: protein-cofactor (quinones and Fe2+) interactions. Proc Natl Acad Sci USA 85: 8487–8491

    Google Scholar 

  • Arata H and Parson WW (1981) Enthalpy and volume changes accompanying electron transfer from P870 to quinones in Rhodopseudomonas sphaeroides reaction centers. Biochim Biophys Acta 636: 70–81

    Google Scholar 

  • Baciou L, Rivas E and Sebban P (1990) P+QA and P+QB charge recombination in Rhodopseudomonas viridis chromatophores and in reaction centers reconstituted in phosphatidylcholine liposomes. Existence of two conformational states of the reaction centers and effects of pH and o-phenanthroline. Biochemistry 29: 2966–2976

    Google Scholar 

  • Van den Brink JS, Hulsebosch RJ, Gast P, Hore PJ and Hoff AJ (1994) QA Binding in reaction centers of the photosynthetic purple bacterium Rhodobacter sphaeroides R26 investigated with electron spin polarization spectroscopy. Biochemistry 33: 13668–13677

    Google Scholar 

  • Brzezinski P and Andréasson L-E (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

    Google Scholar 

  • Brzezinski P, Okamura MY and Feher G (1992) Structural changes following the formation of D+QB in bacterial reaction centers: measurement of light-induced electrogenic events in RCs incorporated in a phospholipid monolayer. In: Breton J and Verméglio A (eds) The Photosynthetic Bacterial Reaction Center II: Structure, Spectroscopy and Dynamics, pp 321–330. Plenum Press, New York and London

    Google Scholar 

  • Calvo R, Butler WF, Isaacson RA, Okamura MY, Fredkin DR and Feher G (1982) Spin-lattice relaxation time of the reduced primary quinone in RCs from R.sphaeroides: Determination of zero-field splitting of Fe2+. Biophys J 37: 111a

    Google Scholar 

  • Calvo R, Passeggi MCG, Isaacson RA, Okamura MY and Feher G (1990) Electron paramagnetic resonance investigation of photosynthetic reaction centers from Rhodobacter sphaeroides R-26 in which Fe2+ was replaced by Cu2+. Determination of hyperfine interactions and exchange and dipole-dipole interaction between Cu2+ and QB. Biophys J 58: 149–165

    Google Scholar 

  • Cohen-Bazire G, Sistrom WR and Stanier RY (1957) Kinetic studies of pigment synthesis by non-sulfur purple bacteria. J Cellular Comp Physiol 49: 25–51, 58–68

    Google Scholar 

  • Debus RJ (1985) The photosynthetic reaction center from Rhodopseudomonas sphaeroides R26.1: Influence of the H subunit and the Fe2+ on electron transfer, pp 59–61. PhD thesis. University of California at San Diego, La Jolla, CA

    Google Scholar 

  • Debus RJ, Feher G and Okamura MY (1985) LM Complex of reaction centers from Rhodopseudomonas sphaeroides R-26: Characterization and reconstitution with the H subunit. Biochemistry 24: 2488–2500

    Google Scholar 

  • Debus RJ, Feher G and Okamura MY (1986) Iron-depleted reaction centers from Rhodopseudomonas sphaeroides R-26.1: Characterization and reconstitution with Fe2+, Mn2+, Co2+, Ni2+, Cu2+, and Zn2+. Biochemistry 25: 2276–2287

    Google Scholar 

  • McElroy JD, Mauzerall DC and Feher G (1974) Characterization of primary reactants in bacterial photosynthesis. II. Kinetic studies of the light-induced EPR signal (g = 2.0026) and the optical absorbance changes at cryogenic temperatures. Biochim Biophys Acta 333: 261–277

    Google Scholar 

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

    Google Scholar 

  • Ermler U, Michel H and Schiffer M (1994b) Structure and function of the photosynthetic reaction center from Rhodobacter sphaeroides. J Bioenerg Biomembr 26: 5–15

    Google Scholar 

  • Feher G and Okamura MY (1978) Chemical composition and properties of reaction centers. In: Sistrom WR and Clayton RK (eds) The Photosynthetic Bacteria, pp 349–386. Plenum Press, New York.

    Google Scholar 

  • Feher G, Isaacson RA, McElroy JD, Ackerson LC and Okamura MY (1974) On the question of the primary acceptor in bacterial photosynthesis: manganese substituting for iron in reaction centers of Rhodopseudomonas spheroides R-26. Biochim Biophys Acta 368: 135–139

    Google Scholar 

  • Feher G, Allen JP, Okamura MY and Rees DC (1989) Structure and function of bacterial photosynthetic reaction centres. Nature 339: 111–116

    Google Scholar 

  • Gao J-L, Shopes RJ and Wraight CA (1991) Heterogeneity of kinetics and electron transfer equilibria in the bacteriopheophytin and quinone electron acceptors of reaction centers from Rhodopseudomonas viridis. Biochim Biophys Acta 1056: 259–272

    Google Scholar 

  • Gunner MR (1991) The reaction center protein from purple bacteria: Structure and function. Current Topics in Bioenergetics 16: 319–367

    Google Scholar 

  • Hienerwadel R, Nabedryk E, Breton J, Kreutz W and Mäntele W (1992) Time-resolved ifnrared and static FTIR studies of QA→QB electron transfer in Rhodopseudomonas viridis reaction centers. In: Breton J and Verméglio A (eds) The Photosynthetic Bacterial Reaction Center II: Structure, Spectroscopy and Dynamics, pp 163–172. Plenum Press, New York

    Google Scholar 

  • Kirmaier C, Holten D and Parson WW (1985) Temperature and detection-wavelength dependence of the picosecond electrontransfer kinetics measured in Rhodopseudomonas sphaeroires reaction centers. Resolution of new spectral and kinetic components in the primary charge-separation process. Biochim Biophys Acta 810: 33–48

    Google Scholar 

  • Kleinfeld D, Okamura MY and Feher G (1984a) Electron-transfer kinetics in photosynthetic reaction centers cooled to cryogenic temperatures in the charge-separated state: Evidence for lightinduced structural changes. Biochemistry 23: 5780–5786

    Google Scholar 

  • Kleinfeld D, Okamura MY and Feher G (1984b) Electron transfer in reaction centers of Rhodopseudomonas sphaeroides. 1. Determination of the charge recombination pathway of D+QAQA and free energy and kinetic relations between QB QB and QAQB. Biochim Biophys Acta 766: 126–140

    Google Scholar 

  • Knox PP, Lukashev EP, Kononenko AA, Venediktov PS and Rubin AB (1977) On possible role of macromolecular components in the functioning of the photosynthetic reaction centres of purple bacteria. Molecular Biology (Moscow) 11: 1090–1099

    Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685

    Google Scholar 

  • Liu B, van Kan PJM and Hoff AJ (1991) Influence of the H-subunit and Fe2+ on electron transport from I? to Q A in Fe2+-free and/or H-free reaction centers from Rhodobacter sphaeroides R-26. FEBS Lett 289: 23–28

    Google Scholar 

  • Malkin S, Churio MS, Shochat S, and Braslavsky SE (1994) Photochemical energy storage and volume changes in the microsecond time range in bacterial photosynthesis – a laser induced optoacoustic study. J Photochem Photobiol B: Biol 23: 79–85

    Google Scholar 

  • Moser CC, Keske JM, Warncke K, Farid RS and Dutton PL (1992) Nature of biological electron transfer. Nature 355: 796–802

    Google Scholar 

  • Nabedryk E, Bagley KA, Thibodeau DL, Bauscher M, Mäntele W and Breton J (1990) A protein conformational change associated with the photoreduction of the primary and secondary quinones in the bacterial reaction center. FEBS Lett 266: 59–62

    Google Scholar 

  • Nam HK, Austin RH and Dismukes GC (1984) On the role of Fe2+ in bacterial photosynthesis. The effect of biosynthetic substitution of Fe2+ by Mn2+ on the electron transfer step Q1 ?Q2 → Q1Q2 ? in reaction centers. Biochim Biophys Acta 765: 301–308

    Google Scholar 

  • Pettersson RP and Wobrauschek P (1995) Total-reflection X-ray fluorescence analysis with an 18 kW rotating anode source – first results. Nucl Instr Methods Phys Res A 355: 665–667

    Google Scholar 

  • Stowell MH, McPhilips TM, Rees DC, Soltis SM, Abresch E and Feher G (1997) Light-induced structural changes in photosynthetic reaction center: implication for mechanism of electronproton transfer. Science 276: 812–816

    Google Scholar 

  • Straley SC, Parson WW, Mauzerall DC and Clayton RK (1973) Pigment content and molar extinction coefficients of photochemical reaction centers from Rhodopseudomonas sphaeroides. Biochim Biophys Acta 305: 597–609

    Google Scholar 

  • Tiede DM and Hanson DK (1992) Protein relaxation following quinone reduction in Rhodobacter capsulatus: Detection of likely protonation-linked optical absorbance changes of the chromatophores. In: Breton J and Verméglio A (eds) The Photosynthetic Bacterial Reaction Center II: Structure, Spectroscopy and Dynamics, pp 341–350. Plenum Press, New York

    Google Scholar 

  • Tiede DM, Vazquez J, Cordova J and Marone PA (1996) Timeresolved electrochromism associated with the formation of quinone anions in the Rhodobacter sphaeroides R26 reaction center. Biochemistry 35: 10763–10775

    Google Scholar 

  • Williams JC, Steiner LA, Feher G (1986) Primary structure of the reaction center from Rhodopseudomonas sphaeroides. Proteins: Structure, Function and Genetics 1: 312–325 Wraight CA (1978) Iron-quinone interactions in the electron acceptor regions of bacterial photosynthetic reaction centers. FEBS Lett 93: 283–288

    Google Scholar 

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Authors and Affiliations

  1. Department of Biochemistry and Biophysics, University of Göteborg, Medicinaregatan 9C, S-41390, Göteborg, Sweden

    I.A. Smirnova, A. Blomberg, L.-E. Andréasson & P. Brzezinski

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  1. I.A. Smirnova
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  2. A. Blomberg
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  3. L.-E. Andréasson
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  4. P. Brzezinski
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Smirnova, I., Blomberg, A., Andréasson, LE. et al. Localization of light-induced structural changes in bacterial photosynthetic reaction centers. Photosynthesis Research 56, 45–55 (1998). https://doi.org/10.1023/A:1005934411312

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