Skip to main content
SpringerLink
Log in

Single Molecule Fluorescence of Native and Refolded Peridinin–Chlorophyll–Protein Complexes

  • Original Paper
  • Published:
Journal of Fluorescence Aims and scope Submit manuscript

Abstract

Single molecule spectroscopy was applied to study the optical properties of native and refolded peridinin–chlorophyll–protein (PCP) complexes. The native system is a trimer with six chlorophyll a (Chl a) molecules, while the refolded one contains two Chl a and resembles structurally and spectroscopically the PCP monomer. The fluorescence emission of single PCP complexes strongly broadens with increasing excitation power. Simultaneously, the distribution of fluorescence maximum frequencies is also broadened. These spectral changes are attributed to photoinduced conformational changes of the protein that influence the fluorescence of embedded chromophores. Comparison of fluorescence intensities measured for PCP complexes in two different solvents indicates that the native PCP trimers are preserved in EDTA Tris buffer, while in PVA polymer matrix only monomers are stable.

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
Fig. 5

Similar content being viewed by others

References

  1. Frauenfelder H (2003) Complexity of proteins. In: Physics of biological systems: from molecules to species. Springer, Heidelberg, Germany

    Google Scholar 

  2. Polivka T, Sundström V (2004) Ultrafast dynamics of carotenoid excited states—from solution to natural and artificial system. Chem Rev 104:2021–2071

    Article  PubMed  CAS  Google Scholar 

  3. Moerner WE (2002) A dozen years of single-molecule spectroscopy in physics, chemistry, and biophysics. J Phys Chem B 106:910–927

    Article  CAS  Google Scholar 

  4. Hofmann C, Aartsma TJ, Michel H, Köhler J (2003) Direct observation of tiers in the energy landscape of a chromoprotein: a single-molecule study. Proc Natl Acad Sci U S A 100:15534–15538

    Article  PubMed  CAS  Google Scholar 

  5. Rutkauskas D, Novoderezhkin V, Cogdell RJ, van Grondelle R (2005) Fluorescence spectroscopy of conformational changes of single LH2 complexes. Biophys J 88:422–435

    Article  PubMed  CAS  Google Scholar 

  6. Valkunas L, Janusonisa J, Rutkauskas D, van Grondelle R (2007) Protein dynamics revealed in the excitonic spectra of single LH2 complexes. J Lumin 127:269–275

    Article  CAS  Google Scholar 

  7. Hofmann E, Wrench PM, Sharples FP, Hiller RG, Welte W, Diederichs K (1996) Structural basis of light harvesting by carotenoids: peridinin chlorophyll-a protein from Amphidinium carterae. Science 272:788–1791

    Article  Google Scholar 

  8. Kleima FJ, Wendling M, Hofmann E, Peterman EJ, van Grondelle R, van Amerongen H (2000) Peridinin chlorophyll a protein: relating structure and steady-state spectroscopy. Biochemistry 39:5184–5195

    Article  PubMed  CAS  Google Scholar 

  9. Wörmke S, Mackowski S, Brotosudarmo THP, Bräuchle C, Garcia A, Braun P, Scheer H, Hofmann E (2007) Detection of single biomolecule fluorescence excited through energy transfer: application to light-harvesting complexes. Appl Phys Lett 90:193901

    Article  CAS  Google Scholar 

  10. Wörmke S, Mackowski S, Brotosudarmo THP, Jung C, Zumbusch A, Ehrl M, Scheer H, Hofmann E, Hiller RG, Bräuchle C (2007) Monitoring fluorescence of individual chromophores in peridinin–chlorophyll–protein complex using single molecule spectroscopy. Biochim Biophys Acta 1767:956–964

    Article  PubMed  CAS  Google Scholar 

  11. Kleima FJ, Hofmann E, Gobets B, Van Stokkum IHM, van Grondelle R, Diederich K, van Amerongen H (2000) Förster excitation energy transfer in peridinin–chlorophyll a–protein. Biophys J 78:344–353

    PubMed  CAS  Google Scholar 

  12. Miller DJ, Catmull J, Puskeiler R, Tweedale H, Sharples FP, Hiller RG (2005) Reconstitution of the peridinin–chlorophyll a protein (PCP): evidence for functional flexibility in chlorophyll binding. Photosynth Res 86:229–240

    Article  PubMed  CAS  Google Scholar 

  13. Brotosudarmo THP, Hofmann E, Hiller RG, Wörmke S, Mackowski S, Zumbusch A, Bräuchle C, Scheer H (2006) Peridinin–chlorophyll–protein reconstituted with chlorophyll mixtures: preparation, bulk and single molecule spectroscopy. FEBS Lett 580:5257–5262

    Article  PubMed  CAS  Google Scholar 

  14. Mackowski S, Wörmke S, Brotosudarmo THP, Jung C, Hiller RG, Scheer H, Bräuchle C (2007) Energy transfer in reconstituted peridinin–chlorophyll–protein complexes: ensemble and single molecule spectroscopy studies. Biophys J 93:3249–3258

    Article  PubMed  CAS  Google Scholar 

  15. Polívka T, Pascher T, Sundström V, Hiller RG (2005) Tuning energy transfer in the peridinin–chlorophyll complex by reconstitution with different chlorophylls. Photosynth Res 86:217–227

    Article  PubMed  CAS  Google Scholar 

  16. Mackowski S, Wörmke S, Brotosudarmo THP, Scheer H, Bräuchle C (2008) Fluorescence spectroscopy of reconstituted peridinin–chlorophyll–protein complexes. Photosynth Res 95:253–260

    Article  PubMed  CAS  Google Scholar 

  17. Law CJ, Cogdell RJ (1998) The effect of chemical oxidation on the fluorescence of the LH1 (B880) complex from the purple bacterium Rhodobium marimum. FEBS Lett 432:27–30

    Article  PubMed  CAS  Google Scholar 

  18. Spezia R, Aschi M, Nola AD, Valentin MD, Carbonera D, Amadei A (2003) The effect of protein conformational flexibility on the electronic properties of a chromophore. Biophys J 84:2805–2813

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgment

The authors thank Eckhard Hofmann for many fruitful discussions. The work was supported by the Deutsche Forschungsgemeinschaft, Bonn (SFB 533, projects A6 and B7) and “Center for Integrated Protein Science Munich (CiPSM)”. S. M. acknowledges financial support from the Alexander von Humboldt Foundation.

Author information

Authors and Affiliations

  1. Department of Chemistry and Biochemistry and Center for Nanoscience, Ludwig-Maximilian-University Munich, 81377, Munich, Germany

    Stephan Wörmke, Andreas Schaller, Tatas H. P. Brotosudarmo & Christoph Bräuchle

  2. Institute of Physics, Nicolaus Copernicus University, Grudziadzka 5, 87-100, Torun, Poland

    Sebastian Mackowski

  3. Department of Biology 1, Ludwig-Maximilians-University, 80638, München, Germany

    Tatas H. P. Brotosudarmo & Hugo Scheer

  4. Department of Biology and Biotechnology, Ruhr-University Bochum, 44780, Bochum, Germany

    Silke Johanning

Authors
  1. Stephan Wörmke
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Sebastian Mackowski
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Andreas Schaller
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Tatas H. P. Brotosudarmo
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Silke Johanning
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Hugo Scheer
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Christoph Bräuchle
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Sebastian Mackowski.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wörmke, S., Mackowski, S., Schaller, A. et al. Single Molecule Fluorescence of Native and Refolded Peridinin–Chlorophyll–Protein Complexes. J Fluoresc 18, 611–617 (2008). https://doi.org/10.1007/s10895-008-0310-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10895-008-0310-9

Keywords

Search

Navigation