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Trapping of a long-living charge separated state of photosynthetic reaction centers in proteoliposomes of negatively charged phospholipids

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Abstract

Reaction centers from the purple bacterium Rhodobacter sphaeroides strain R-26.1 were purified and reconstituted in proteoliposomes formed by the anionic phospholipids phosphatidylglycerol, phosphatidylserine and phosphatidylinositol and by the zwitterionic phospholipid phosphatidylcholine by size-exclusion chromatography in the dark and under illumination. We report the large stabilizing effect induced by anionic phospholipids on the protein charge separated state which results trapped in a long-living (up to tens of minutes) state with a yield up to 80%. This fully reversible state is formed in oxygenic conditions regardless the presence of the secondary quinone QB and its lifetime and relative yield increase at low pH. In proteoliposomes formed with QA-depleted reaction centers (RCs) the resulting protein is very light-sensitive and the long living charge separated state is not observed. The data collected in negatively charged proteoliposomes are discussed in terms of the electrostatic effect on the primary quinone acceptor and compared with similar long living species reported in literature and obtained in anionic, zwitterionic, and non-ionic detergents.

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References

  • U Andréasson L-E Andréasson (2003) ArticleTitleCharacterization of a semi-stable, charge-separated state in reaction centers from Rhodobacter sphaeroides Photosynth Res 75 IssueID3 223–233 Occurrence Handle10.1023/A:1023944605460

    Article  Google Scholar 

  • U Andréasson T Carlsson L-E Andréasson (2003) ArticleTitleSpectroscopic characterization of a semi-stable, charge-separated state in Cu2+ substituted reaction centers from Rhodobacter sphaeroides Biochim et Biophysica Acta (BBA)–Bioenergetics 1607 IssueID3 45–52 Occurrence Handle10.1016/j.bbabio.2003.08.007

    Article  Google Scholar 

  • KM Andrews AR Crofts RB Gennis (1990) ArticleTitleLarge-scale purification and characterization of a highly active four- subunit cytochrome bc1 complex from Rhodobacter sphaeroides Biochemistry 29 IssueID11 2645–2651 Occurrence Handle10.1021/bi00463a004 Occurrence Handle2161250

    Article  PubMed  Google Scholar 

  • A Camara-Artigas D Brune JP Allen (2002) ArticleTitleInteractions between lipids and bacterial reaction centers determined by protein crystallography Proc Natl Acad Sci USA 99 IssueID17 11055–11060 Occurrence Handle12167672

    PubMed  Google Scholar 

  • MS el Kebbaj N Latruffe (1986) ArticleTitleKinetic aspects of the role of phospholipids in D-beta-hydroxybutyrate dehydrogenase activity Arch Biochem Biophys 244 IssueID2 662–670 Occurrence Handle10.1016/0003-9861(86)90634-X Occurrence Handle3947085

    Article  PubMed  Google Scholar 

  • MS el Kebbaj N Latruffe M Monsigny A Obrenovitch (1986) ArticleTitleInteractions between apo-(D-beta-hydroxybutyrate dehydrogenase) and phospholipids studied by intrinsic and extrinsic fluorescence Biochem J 237 IssueID2 359–364 Occurrence Handle3800892

    PubMed  Google Scholar 

  • I Fathir T Mori T Nogi M Kobayashi K Miki T Nozawa (2001) ArticleTitleStructure of the H subunit of the photosynthetic reaction center from the thermophilic purple sulfur bacterium, Thermochromatium tepidum Implications for the specific binding of the lipid molecule to the membrane protein complex Eur J Biochem 268 IssueID9 2652–2657 Occurrence Handle10.1046/j.1432-1327.2001.02158.x Occurrence Handle11322886

    Article  PubMed  Google Scholar 

  • G Feher (1971) ArticleTitleSome chemical and physical properties of a bacterial reaction center particle and its primary photochemical reactants Photochem Photobiol 14 IssueID3 373–388 Occurrence Handle4330229

    PubMed  Google Scholar 

  • PK Fyfe NW Isaacs RJ Cogdell MR Jones (2004) ArticleTitleDisruption of a specific molecular interaction with a bound lipid affects the thermal stability of the purple bacterial reaction centre Biochim Biophys Acta Bioenerg 1608 IssueID1 11–22 Occurrence Handle10.1016/j.bbabio.2003.09.014

    Article  Google Scholar 

  • RB Gennis (1989) Biomembranes: Molecular Structure and Function Springer-Verlag New York

    Google Scholar 

  • RB Gennis A Jonas (1977) ArticleTitleProtein lipid interactions Annu Revi Biophys Bioenerg 6 195–238 Occurrence Handle10.1146/annurev.bb.06.060177.001211

    Article  Google Scholar 

  • AO Goushcha VN Kharkyanen GW Scott AR Holzwarth (2000) ArticleTitleSelf-regulation phenomena in bacterial reaction centers I. General theory. Biophys J 79 IssueID3 1237–1252

    Google Scholar 

  • AO Goushcha AJ Manzo et al. (2003) ArticleTitleSelf-regulation phenomena applied to bacterial reaction centers: 2 Nonequilibrium adiabatic potential: dark and light conformations revisited Biophys J 84(2 Pt 1) 1146–1160

    Google Scholar 

  • AO Goushcha AJ Manzo VN Kharkyanen R Grondelle Particlevan GW Scott (2004) ArticleTitleLight-induced equilibration kinetics in membrane-bound photosynthetic reaction centers: nonlinear dynamic effects in multiple scattering media J Phys Chem B 108 IssueID8 2717–2725 Occurrence Handle10.1021/jp036403d

    Article  Google Scholar 

  • JF Imhoff U Bias-Imhoff (1995) Lipids and quinones RE Blankenship TM Madigan CE Bauer (Eds) Anoxygenic Photosynthetic Bacteria Kluwer Academic Publishers Dordrecht, The Netherlands 179–205

    Google Scholar 

  • JF Imhoff DJ Kushner SC Kushwaha M Kates (1982) ArticleTitlePolar lipids in phototrophic bacteria of the Rhodospirillaceae and Chromatiaceae families J Bacteriol 150 IssueID3 1192–1201 Occurrence Handle7076618

    PubMed  Google Scholar 

  • RA Isaacson F Lendzian EC Abresch W Lubitz G Feher (1995) ArticleTitleElectronic structure of QA − in reaction centers from Rhodobacter sphaeroides I. Electron paramagnetic resonance in single crystals Biophys J 69 IssueID2 311–322 Occurrence Handle8527644

    PubMed  Google Scholar 

  • L Kálmán P Maróti (1997) ArticleTitleConformation-Activated Protonation in Reaction Centers of the Photosynthetic Bacterium Rhodobacter sphaeroides Biochemistry 36 IssueID49 15269–15276 Occurrence Handle10.1021/bi971882q Occurrence Handle9398255

    Article  PubMed  Google Scholar 

  • D Kleinfeld MY Okamura G Feher (1984) ArticleTitleElectron transfer in reaction centres of Rhodopseudomonas sphaeroides I. Determination of the charge recombination pathway of QA −QB and free energy and kinetic relations between QA −QB and QA −QB − Biochim Biophys Acta 766 126–140 Occurrence Handle6331502

    PubMed  Google Scholar 

  • Kutuzov MA (1990). Structure-functional studies of the photosynthetic reaction centre of the green thermophilic bacterium Chloroflexus aurantiacus: primary structure, topography and mechanisms of thermal stability. Shemyakin Institute of Bioorganic Chemistry. Moscow, Moscow State University.

  • MA Kutuzov MD Mamedov A Semenov VP Shinkarev MI Verkhovsky NG Abdulaev LA Drachev (1991) ArticleTitleFunctioning of quinone acceptors in the reaction center of the green photosynthetic bacterium Chloroflexus aurantiacus FEBS Lett 289 IssueID2 179–82 Occurrence Handle10.1016/0014-5793(91)81064-F Occurrence Handle1915846

    Article  PubMed  Google Scholar 

  • N Latruffe JM Berrez MS el Kebbaj (1986) ArticleTitleLipid-protein interactions in biomembranes studied through the phospholipid specificity of D-beta-hydroxybutyrate dehydrogenase Biochimie 68 IssueID3 481–491 Occurrence Handle3091085

    PubMed  Google Scholar 

  • KE McAuley PK Fyfe JP Ridge NW Isaacs RJ Cogdell MR Jones (1999) ArticleTitleStructural details of an interaction between cardiolipin and an integral membrane protein Proc Natl Acad Sci USA 96 IssueID26 14706–14711 Occurrence Handle10611277

    PubMed  Google Scholar 

  • F Milano A Agostiano F Mavelli M Trotta (2003) ArticleTitleKinetics of the Quinone Binding reaction at the QB site of Reaction centers from the purple bacteria Rhodobacter sphaeroides reconstituted in liposomes Eur J Biochem 270 4595–4605 Occurrence Handle10.1046/j.1432-1033.2003.03845.x Occurrence Handle14622246

    Article  PubMed  Google Scholar 

  • F Mueh J Rautter W Lubitz (1997) ArticleTitleTwo distinct conformations of the primary electron donor in reaction centres from Rhodobacter sphaeroides revealed by ENDOR/TRIPLE-spectroscopy Biochemistry 36 4155–4162 Occurrence Handle10.1021/bi962859s Occurrence Handle9100009

    Article  PubMed  Google Scholar 

  • L Nagy E Fodor T Farkas S Gedey A Kecskes (1998) Lipids effect the charge stabilisation in wild type and mutant reaction centres of photosynthetic bacteria Rhodobacter sphaeroides G Garab (Eds) Photosynthesis: Mech Eff. Proc. Int. Congr. Photosynth., 11th Kluwer Academic Publishers Dordrecht, The Netherlands 893–896

    Google Scholar 

  • L Nagy E Fodor T Farkas S Gedey A Kecskes (1999) ArticleTitleLipids effect the charge stabilisation in wild type and mutant reaction centres of photosynthetic bacteria Rhodobacter sphaeroides Aust J Plant Physiol 25 465–473

    Google Scholar 

  • MY Okamura RA Isaacson G Feher (1975) ArticleTitleThe primary acceptor in bacterial photosynthesis: the obligatory role of ubiquinone in photoactive reaction centres of Rp sphaeroides Proc Natl Acad Sci USA 72 3491–3495 Occurrence Handle1081231

    PubMed  Google Scholar 

  • RE Overfield CA Wraight (1980) ArticleTitleOxidation of cytochromes c and c2 by bacterial photosynthetic reaction centers in phospholipid vesicles 1. Studies with neutral membranes Biochemistry 19 IssueID14 3322–3327 Occurrence Handle10.1021/bi00555a034 Occurrence Handle6250565

    Article  PubMed  Google Scholar 

  • G Palazzo A Mallardi M Giustini D Berti G Venturoli (2000) ArticleTitleCumulant analysis of charge recombination kinetics in bacterial reaction centers reconstituted into lipid vesicles Biophys J 79 IssueID3 1171–1179 Occurrence Handle10968981

    PubMed  Google Scholar 

  • L Rinyu EW Martin E Takahashi P Maroti C Wraight (2004) ArticleTitleModulation of the free energy of the primary quinone acceptor (QA) in Reaction Centers from Rhodobacter sphaeroides: Contribution from the protein and the protein-lipid (cardiolipin) interactions Biochim Biophys Acta Bioenerg 1655 93–101 Occurrence Handle10.1016/j.bbabio.2003.07.012

    Article  Google Scholar 

  • H Sandermann SuffixJr (1978) ArticleTitleRegulation of membrane enzymes by lipids Biochim Biophys Acta 515 IssueID3 209–237 Occurrence Handle359048

    PubMed  Google Scholar 

  • H Sandermann SuffixJr (1982) ArticleTitleLipid-dependent membrane enzymes A kinetic model for cooperative activation in the absence of cooperativity in lipid binding Eur J Biochem 127 IssueID1 123–128 Occurrence Handle10.1111/j.1432-1033.1982.tb06845.x Occurrence Handle7140750

    Article  PubMed  Google Scholar 

  • H Sandermann SuffixJr (1983) ArticleTitleLipid Solvation and Kinet Cooperativity of Function Membrane proteins Trends Biol Sci 8 408–411 Occurrence Handle10.1016/0968-0004(83)90306-7

    Article  Google Scholar 

  • H Sandermann (2002) ArticleTitleHigh free energy of lipid/protein interaction in biological membranes FEBS Lett 514 IssueID2-3 340–342 Occurrence Handle10.1016/S0014-5793(02)02400-6 Occurrence Handle11943177

    Article  PubMed  Google Scholar 

  • M Trotta F Milano L Nagy A Agostiano (2002) ArticleTitleResponse of membrane protein to the environment: the case of photosynthetic reaction centre Mater Sci Eng C 22 IssueID2 263–267 Occurrence Handle10.1016/S0928-4931(02)00178-9

    Article  Google Scholar 

  • M RestVan ParticleVan der G Gingras (1974) ArticleTitleThe Pigment Complement of the Photosynthetic Reaction Center Isolated from Rhodospirillum rubrum J Biol Chem 249 IssueID20 6446–6453 Occurrence Handle4214257

    PubMed  Google Scholar 

  • F Mourik Particlevan M Reus AR Holzwarth (2001) ArticleTitleLong-lived charge-separated states in bacterial reaction centers isolated from Rhodobacter sphaeroides Biochim Biophys Acta 1504 IssueID2-3 311–318 Occurrence Handle11245794

    PubMed  Google Scholar 

  • G Venturoli M Trotta R Feick BA Melandri D Zannoni (1991) ArticleTitleTemperature dependence of charge recombination from the P+QA + abd P+QB − states in photosynthetic reaction centres isolated from thermophilic bacterium Chloroflexus aurantiacus Eur J Biochem 202 625–634 Occurrence Handle10.1111/j.1432-1033.1991.tb16416.x Occurrence Handle1761060

    Article  PubMed  Google Scholar 

  • JC Williams ALM Haffa JL McCulley NW Woodbury JP Allen (2001) ArticleTitleElectrostatic interactions between charged amino acid residues and the bacteriochlorophyll dimer in reaction centers from Rhodobacter sphaeroides Biochemistry 40 15403–15407 Occurrence Handle10.1021/bi011574z Occurrence Handle11735424

    Article  PubMed  Google Scholar 

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Authors and Affiliations

  1. Istituto per i Processi Chimico-Fisici, Sezione di Bari, Università di Bari, Via Orabona 4, Bari, 70126, Italy

    Angela Agostiano, Francesco Milano & Massimo Trotta

  2. Dipartimento di Chimica, Università di Bari, Via Orabona 4, Bari, 70126, Italy

    Angela Agostiano

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  1. Angela Agostiano
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  2. Francesco Milano
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  3. Massimo Trotta
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Correspondence to Massimo Trotta.

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Agostiano, A., Milano, F. & Trotta, M. Trapping of a long-living charge separated state of photosynthetic reaction centers in proteoliposomes of negatively charged phospholipids. Photosynth Res 83, 53–61 (2005). https://doi.org/10.1007/s11120-004-3197-6

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  • DOI: https://doi.org/10.1007/s11120-004-3197-6

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