Skip to main content
SpringerLink
Log in

Comparison of the fluorescence kinetics of detergent-solubilized and membrane-reconstituted LH2 complexes from Rps. acidophila and Rb. sphaeroides

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

Abstract

Picosecond time-resolved fluorescence spectroscopy has been used in order to compare the fluorescence kinetics of detergent-solubilized and membrane-reconstituted light-harvesting 2 (LH2) complexes from the purple bacteria Rhodopseudomonas (Rps.) acidophila and Rhodobacter (Rb.) sphaeroides. LH2 complexes were reconstituted in phospholipid model membranes at different lipid:protein-ratios and all samples were studied exciting with a wide range of excitation densities. While the detergent-solubilized LH2 complexes from Rps. acidophila showed monoexponential decay kinetics (τf = 980 ps) for excitation densities of up to 3·1013 photons/(pulse·cm2), the membrane-reconstituted LH2 complexes showed multiexponential kinetics even at low excitation densities and high lipid:protein-ratios. The latter finding indicates an efficient clustering of LH2 complexes in the phospholipid membranes. Similar results were obtained for the LH2 complexes from Rb. sphaeroides.

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Abbreviations

BChl:

Bacteriochlorophyll

β-OG:

Octyl-β-d-glucopyranoside

DOPC:

1,2-Dioleoyl-sn-glycero-3-phosphocholine

DOPG:

1,2-Dioleoyl-sn-glycero-3-phosphoglycerol

f:

Focal length

GlyGly:

Glycyl–glycine buffer

IRF:

Instrument response function

LDAO:

Lauryldimethylamine N-oxide

LH2:

Light-harvesting 2 complex

L:P:

Lipid:protein-ratio

Rb.:

Rhodobacter

Rps.:

Rhodopseudomonas

Tris:

Tris-hydroxymethyl-aminomethane

τ:

Fluorescence lifetime

References

  • Bopp MA, Jia Y, Li L, Cogdell RJ, Hochstrasser RM (1997) Fluorescence and photobleaching dynamics of single light-harvesting complexes. Proc Natl Acad Sci USA 94:10630–10635

    Article  PubMed  CAS  Google Scholar 

  • Chen X-H, Zhang L, Weng Y-X, Du L-C, Ye M-P, Yang G-Z, Fujii R, Rondonuwu FS, Koyama Y, Wu Y-S, Zhang JP (2005) Protein structural deformation induced lifetime shortening of photosynthetic bacteria light-harvesting complex LH2 excited states. Biophys J 88:4262–4273

    Article  PubMed  CAS  Google Scholar 

  • Cogdell RJ, Durant I, Valentine J, Lindsay JG, Schmidt K (1983) The isolation and partial characterisation of the light-harvesting pigment-protein complement of Rhodopseudomonas acidophila. Biochim Biophys Acta 722:427–435

    Article  CAS  Google Scholar 

  • Cogdell RJ, Gall A, Köhler J (2006) The architecture and function of purple bacteria: from single molecules to in vivo membranes. Q Rev Biophys 39:227–324

    Article  PubMed  CAS  Google Scholar 

  • de Ruijter WPF, Oellerich S, Segura J-M, Lawless AM, Papiz M, Aartsma TJ (2004) Observation of the energy-level structure of the low-light adapted B800 LH4 complex by single-molecule spectroscopy. Biophys J 87:3413–3420

    Article  PubMed  CAS  Google Scholar 

  • Francia F, Wang J, Venturoli G, Melandri BA, Barz WP, Oesterhelt D (1999) The reaction center-LH1 antenna complex of Rhodobacter sphaeroides contains one PufX molecule which is involved in dimerization of this complex. Biochemistry 38:6834–6845

    Article  PubMed  CAS  Google Scholar 

  • Francia F, Dezi M, Rebechi A, Mallardi A, Palazzo G, Melandri BA, Venturoli G (2004) Light-harvesting complex 1 stabilizes P+Q −B charge separation in reaction centers of Rhodobacter sphaeroides. Biochemistry 43:14199–14210

    Article  PubMed  CAS  Google Scholar 

  • Freiberg A, Timpmann K, Lin S, Woodbury NW (1998) Exciton relaxation and transfer in the LH2 antenna network of photosynthetic bacteria. J Phys Chem B 102:10974–10982

    Article  CAS  Google Scholar 

  • Freiberg A, Rätsep M, Timpmann K, Trinkunas G, Woodbury NW (2003) Self-trapped excitons in LH2 antenna complexes between 5 K and ambient temperature. J Phys Chem B 107:11510–11519

    Article  CAS  Google Scholar 

  • Freiberg A, Rätsep M, Timpmann K, Trinkunas G (2004) Dual fluorescence of single LH2 antenna nanorings. J Lumin 108:107–110

    Article  CAS  Google Scholar 

  • Gardiner AT, Cogdell RJ, Takaichi S (1993) The effect of growth conditions on the light-harvesting apparatus in Rhodopseudomonas acidophila. Photosynth Res 38:159–167

    Article  CAS  Google Scholar 

  • Gonçalves RP, Busselez J, Lévy D, Seguin J, Scheuring S (2005) Membrane insertion of Rhodopseudomonas acidophila light harvesting complex 2 investigated by high resolution AFM. J Struct Biol 149:79–86

    Article  PubMed  CAS  Google Scholar 

  • Hess S, Chachisvilis M, Timpmann K, Jones M, Fowler G, Hunter C, Sundström V (1995) Temporally and spectrally resolved subpicosecond energy transfer within the peripheral antenna complex (LH2) and from LH2 to the core antenna complex in photosynthetic purple bacteria. Proc Natl Acad Sci USA 92:12333–12337

    Article  PubMed  CAS  Google Scholar 

  • Hofmann C, Aartsma TJ, Köhler J (2004) Energetic disorder and the B850-exciton states of individual light-harvesting 2 complexes from Rhodopseudomonas acidophila. Chem Phys Lettt 395:373–378

    Article  CAS  Google Scholar 

  • Hunter CN, Bergström H, van Grondelle R, Sundstsröm V (1990) Energy-transfer dynamics in three light-harvesting mutants of Rhodobacter sphaeroides: a picosecond spectroscopy study. Biochemistry 29:3203–3207

    Article  PubMed  CAS  Google Scholar 

  • Jimenez R, Dikshit SN, Bradford SE, Fleming GR (1996) Electronic excitation transfer in the LH2 complex of Rhodobacter sphaeroides. J Phys Chem 100:6825–6834

    Article  CAS  Google Scholar 

  • McDermott G, Prince SM, Freer AA, Hawthornthwaite-Lawless AM, Papiz MZ, Cogdell RJ, Isaacs NW (1995) Crystal structure of an integral membrane light-harvesting complex from photosynthetic bacteria. Nature 374:517–521

    Article  CAS  Google Scholar 

  • Milhiet P-E, Gubellini F, Berquand A, Dosset P, Rigaud J-L, Le Grimellec C, Lévy D ( 2006) High-resolution AFM of membrane proteins directly incorporated at high density in planar lipid bilayer. Biophys J 91:3268–3275

    Article  PubMed  CAS  Google Scholar 

  • Monshouwer R, Abrahamsson M, van Mourik F, van Grondelle R (1997) Superradiance and exciton delocalization in bacterial photosynthetic light-harvesting systems. J Phys Chem B 101:7241–7248

    Article  CAS  Google Scholar 

  • Papiz MZ, Prince SM, Howard TD, Cogdell RJ, Isaacs NW (2003) The structure and thermal motion of the B800–850 LH2 complex from Rps. acidophila at 2.0 Å resolution and 100 K: new structural features and functionally relevant motions. J Mol Biol 326:1523–1538

    Article  PubMed  CAS  Google Scholar 

  • Pullerits T, Chachisvilis M, Jones MR, Hunter CN, Sundström V (1994) Exciton dynamics in the light-harvesting complexes of Rhodobacter sphaeroides. Chem Phys Lettt 224:355–365

    Article  CAS  Google Scholar 

  • Richter MF, Baier J, Prem T, Oellerich S, Francia F, Venturoli G, Oesterhelt D, Southall J, Cogdell RJ, Köhler J (2007a) Symmetry matters for the electronic structure of core complexes from Rhodopseudomonas palustris and Rhodobacter sphaeroides PufX-. Proc Natl Acad Sci USA 104:6661–6665

    Article  PubMed  CAS  Google Scholar 

  • Richter MF, Baier J, Cogdell RJ, Köhler J, Oellerich S (2007b) Single-molecule spectroscopic characterization of light-harvesting 2 complexes reconstituted into model membranes. Biophys J 93:183–191

    Article  PubMed  CAS  Google Scholar 

  • Rutkauskas D, Novoderezkhin R, Cogdell RJ, van Grondelle R (2004) Fluorescence spectral fluctuations of single LH2 complexes from Rhodopseudomonas acidophila strain 10050. Biochemistry 43:4431–4438

    Article  PubMed  CAS  Google Scholar 

  • Scheuring S, Gonçalves RP, Prima V, Sturgis JN (2006) The photosynthetic apparatus of Rhodopseudomonas palustris: structures and organization. J Mol Biol 358:83–96

    Article  PubMed  CAS  Google Scholar 

  • Schubert A, Stenstam A, Beenken WJD, Herek JL, Cogdell R, Pullerits T, Sundström V (2004) In vitro self-assembly of the light harvesting pigment-protein LH2 revealed by ultrafast spectroscopy and electron microscopy. Biophys J 86:2363–2373

    PubMed  CAS  Google Scholar 

  • Sebban P, Jolchine G, Moya I (1984) Spectra of fluorescence lifetime and intensity of Rhodopseudomonas sphaeroides at room and low temperature. Comparison between the wild type, the C71 reaction center-less mutant and the B800–850 pigment-protein complex. Photochem Photobiol 39:247–253

    Google Scholar 

  • Stamouli A, Kafi S, Klein DCG, Oosterkamp TH, Frenken JWM, Cogdell RJ, Aartsma TJ (2003) The ring structure and organization of light-harvesting 2 complexes in a reconstituted lipid bilayer, resolved by atomic force microscopy. Biophys J 84:2483–2491

    Article  PubMed  CAS  Google Scholar 

  • Stiel H, Leupold D, Teuchner K, Nowak F, Scheer H, Cogdell RJ (1997) One- and two-exciton bands in the LH2 antenna of Rhodopseudomonas acidophila. Chem Phys Lettt 276:62–69

    Article  CAS  Google Scholar 

  • Sturgis JN, Robert B (1997) Pigment binding-site and electronic properties in light-harvesting proteins of purple bacteria. J Phys Chem B 101:7227–7231

    Article  CAS  Google Scholar 

  • Sundström V, Pullerits T, van Grondelle R (1999) Photosynthetic light-harvesting: reconciling dynamics and structure of purple bacterial LH2 reveals function of photosynthetic unit. J Phys Chem B 103:2327–2346

    Article  Google Scholar 

  • Tietz C, Chekhlov O, Wrachtrup J, Schuster J (1999) Spectroscopy on single light-harvesting complexes at low temperatures. J Phys Chem B 103:6328–6333

    Article  CAS  Google Scholar 

  • van Grondelle R, Hunter CN, Bakker JGC, Kramer HJM (1983) Size and structure of antenna complexes of photosynthetic bacteria as studied by singlet-singlet quenching of the bacteriochlorophyll fluorescence yield. Biochim Biophys Acta 723:30–36

    Article  Google Scholar 

  • van Oijen AM, Ketelaars M, Köhler J, Aartsma TJ, Schmidt J (1999) Unraveling the electronic structure of individual photosynthetic pigment-protein complexes. Science 285:400–402

    Article  PubMed  Google Scholar 

  • Vos M, van Grondelle R, van der Kooij FW, van de Poll D, Amesz J, Duysens LNM (1986) Singlet-singlet annihilation at low temperatures in the antenna of purple bacteria. Biochim Biophys Acta 850:501–512

    Article  CAS  Google Scholar 

  • Vos M, van Dorssen RJ, Amesz J, van Grondelle R, Hunter CN (1988) The organization of the photosynthetic apparatus of Rhodobacter sphaeroides: studies of antenna mutants using singlet-singlet quenching. Biochim Biophys Acta 933:132–140

    Article  CAS  Google Scholar 

  • Vulto SIE, Kennis JTM, Streltsov AM, Amesz J, Aartsma TJ (1999) Energy relaxation within the B850 absorption band of the isolated light-harvesting complex LH2 from Rhodopseudomonas acidophila at low temperature. J Phys Chem B 103:878–883

    Article  CAS  Google Scholar 

  • Westerhuis WHJ, Vos M, van Grondelle R, Amesz J, Niederman RA (1998) Altered organization of light-harvesting complexes in phospholipid-enriched Rhodobacter sphaeroides chromatophores as determined by fluorescence yield and singlet-singlet annihilation measurements. Biochim Biophys Acta 1366:317–329

    Article  CAS  Google Scholar 

Download references

Acknowledgements

T.P. acknowledges financial support from the ‘Bayerische Forschungsstiftung’ (BFS) and R.J.C. thanks the BBSRC for financial support.

Author information

Authors and Affiliations

  1. Lehrstuhl für Experimentalphysik IV, Universität Bayreuth, Universtitätsstrasse 30, 95440, Bayreuth, Germany

    Tobias Pflock, Jürgen Köhler & Silke Oellerich

  2. Department of Biology, University of Bologna and INFM, Via Irnerio 42, Bologna, 40126, Italy

    Manuela Dezi & Giovanni Venturoli

  3. Division of Biochemistry and Molecular Biology, Institute of Biomedical and Life Sciences, University of Glasgow, Glasgow, G12 8QQ, UK

    Richard J. Cogdell

Authors
  1. Tobias Pflock
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Manuela Dezi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Giovanni Venturoli
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Richard J. Cogdell
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Jürgen Köhler
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Silke Oellerich
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Silke Oellerich.

Additional information

Guest editor: Dr. Conrad Mullineaux.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Pflock, T., Dezi, M., Venturoli, G. et al. Comparison of the fluorescence kinetics of detergent-solubilized and membrane-reconstituted LH2 complexes from Rps. acidophila and Rb. sphaeroides . Photosynth Res 95, 291–298 (2008). https://doi.org/10.1007/s11120-007-9245-2

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11120-007-9245-2

Keywords

Search

Navigation