Skip to main content
SpringerLink
Log in

Polarized fluorescence spectroscopy of oriented isolated spinach Photosystem I particles

  • Published:
Photosynthesis Research Aims and scope Submit manuscript

Abstract

The fluorescence anisotropy of Photosystem I (PS I) particles, isolated from spinach chloroplasts and containing approximately 200 chlorophyll molecules per reaction center, is investigated at low temperatures. The particles are oriented by squeezing in polyacrylamid gels with different macroscopic deformation parameters. Fluorescence anisotropy is measured upon steady state excitation with a laser line at 632.8 nm. A formula for the fluorescence anisotropy in oriented Photosystem I particles is applied for a different polarization of the linearly polarized exciting light. Our calculations are based on the consideration of the Photosystem I complex as a triple-chromophore complex: the absorbing chlorophyll molecules (chl), belonging to the light-harvesting complex of PS I (LHC), and two fluorophores, emitting at 720 nm (F720) and at 735 nm (F735), respectively. Using polarized fluorescence spectroscopy with a different polarization of the linearly polarized exciting light, the experimental dependence of the fluorescence anisotropy on this polarization is obtained. Based on this dependence and applying the derived formula, as a first approximation, both the orientation of the photosynthetic pigments with respect to the membrane and their mutual orientation are determined in PS I particles. As the most probable average orientational angles in PS I particles, we obtained the values 35°÷ 50°, 50°÷ 60°, and 65°÷ 67° for the absorbing dipoles of chl and for the emission dipoles of F720 and F735, respectively, with the normal of the plane of the membrane. For their mutual orientation, the following limits are determined: 10°÷ 20°, 40 ± 2°, 20°÷ 30° for the angles between chl and F720, chl and F735; and F720 and F735, correspondingly. Of course, the values of the angles estimated as a result of our study are an average value of all angles of the excited transitions and must be considered as their first approximation valid for the idealized case when all PS I particles are oriented in gel.

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

  • Abdourakhmanov IA, E. Solov'ev AOYu and Chugunov VA (1979) Orientation and linear dichroism of the reaction centres from Rhodopseudomonas sphaeroides R-26. Biochim Biophys Acta 546: 183–186

    Article  PubMed  CAS  Google Scholar 

  • Andreeva A (1996) The rate of energy transfer between long wavelength pigments in isolated Photosystem I particles. In: Marshall JM, Kirov N and Vavrek A (eds) Future Directions in Thin Films — Science and Technology, pp 544–547, World Sci Publ Corp, London

    Google Scholar 

  • Andreeva A and Velitchkova M (1998a) Mutual orientation of absorbing chromophores and long wavelength pigments in Photosystem I particles. Spectrochim Acta Part A 54: 639–644

    Article  Google Scholar 

  • Andreeva A and Velitchkova M (1998b) Orientation of optical transitions of pigments in isolated spinach Photosystem I particles. In: Akos G, Lupkovics G and Podmaniczky A (eds) Optika '98: 5th Congress on Modern Optics, Proceedings of SPIE — The International Society for Optical Engineering 3573: 243–246

  • Beddard GS (1998) Excitation and excitons in Photosystem I. Phil Trans R Soc London A 356: 421–448

    Article  CAS  Google Scholar 

  • Becker JF, Breton JN, Geacintov E and Trentacosti F (1976) Anisotropy of photosynthetic membranes and the degree of fluorescence polarization. Biochim Biophys Acta 440: 531–544

    Article  PubMed  CAS  Google Scholar 

  • Boekema J, Wynn RM and Malkin R (1990) The structure of spinach Photosystem I studied by electron microscopy. Biochim Biophys Acta 1017: 49–56

    Article  CAS  Google Scholar 

  • Breton J and Ikegami I (1989) Orientation of Photosystem-I pigments: low temperature linear dichroism spectroscopy of a highly-enriched P700 particles isolated from spinach. Photosynth Res 21: 27–36

    CAS  Google Scholar 

  • Breton J and Vermeglio A (1982) Orientation of photosynthetic pigments in vivo. In: Govindjee (ed) Photosynthesis: Energy Conversion by Plants and Bacteria, pp 153–194. Academic Press, New York

    Google Scholar 

  • Brettel K (1997) Electron transfer and arrangement of redox cofactors in Photosystem I. Biochim Biophys Acta 13–18: 322–373

    Google Scholar 

  • Demidov A (1994) Fluorescence polarization of triple-chromophore complexes with energy transfer. Applied Optics 33: 6303–6306

    CAS  Google Scholar 

  • Demidov A and Andrews DL (1996) Determination of fluorescence polarization and absorption anisotrepy in molecular complexes having threefold rotational symmetry. Photochem Photobiol 63: 39–52

    CAS  Google Scholar 

  • Fromme P, Witt HT, Schubert WD, Klukas O, Saenger W and, Krauß N (1996) Structure of Photosystem I at 4.5 Å resolution: a short review including evolutionary aspects. Biochim Biophys Acta 1275: 76–83

    Article  Google Scholar 

  • Garab GI and Breton J (1976) Polarized light spectroscopy on oriented spinach chloroplasts. Fluorescence emission at low temperature. Biochem Biophys Res Com 71: 1095–1102

    Article  PubMed  CAS  Google Scholar 

  • Gobets BH, van Amerongen H, Monshouwer R, Kruip J, Rogner M, van Grondellee R and Dekker JP (1994) Polarized site-selected fluorescence spectroscopy of isolated Photosystem I particles, Biochim Biophys Acta 1188: 75–85

    Article  CAS  Google Scholar 

  • Golbeck JH and Bryant DA (1991) Photosystem I. In: Lee CP (ed) Current Topics in Bioenergetics, Vol 16, pp 83–110. Academic Press, New York

    Google Scholar 

  • Haworth P, Arntzen CJ, Tapie P and Breton J (1982) Orientation of pigments in the thylakoid membrane and in isolated chlorophyll-protein complexes of higher plants. I. Determination of optimal conditions for linear dichroism measurements. Biochim Biophys Acta 679: 428–435

    Article  CAS  Google Scholar 

  • Johansson LB-Å and Lindblom G (1980) Orientation and mobility of molecules in membranes studied by polarized light spectroscopy. Q Rev Biophys 13: 63–118

    Article  PubMed  CAS  Google Scholar 

  • Kramer HJM and Amesz J (1982) Anisotropy of the emission and absorption bands of spinach chloroplasts measured by fluorescence polarization and polarized excitation spectra at low temperature. Biochim Biophys Acta 682: 201–207

    Article  Google Scholar 

  • Kiss LI, Ganago AO and Garab GyI (1985) Quantitative method for studying orientation of transition dipoles in membrane vesicles of spherical symmetry. J Biochem Biophys Methods 11: 2113–225

    Article  Google Scholar 

  • Krauss N, Hinrichs W, Witt I, Fromme P, Pritzlow W, Dauter Z, Betzel C, Wilson KS and Saenger W (1993) Three-dimensional structure of system I of photosynthesis at 6 Å resolution. Nature 361: 326–331

    Article  CAS  Google Scholar 

  • Kumazaki S, Ikegami I and Yoshihara K (1997) Excitation and electron transfer from selectively excited primary donor chlorophyll (P700) in a Photosystem I reaction center. J Phys Chem 101: 597–604

    CAS  Google Scholar 

  • Lakowicz JR (1983) Principles of Fluorescence Spectroscopy. Plenum Press, New York

    Google Scholar 

  • Lichtenhaler HK (1987) Chlorophyll and carotenoids: Pigments of photosynthetic biomembranes. Methods Enzymol 148: 350–382

    Article  Google Scholar 

  • Mukerji I and Sauer K (1989) Temperature-dependent steady-state and picosecond kinetic fluorescence measurements of a Photosystem I preparation from spinach. In: Biggins WH (ed) Photosynthesis, pp 105–122, AR Liss, New York

    Google Scholar 

  • Mukkerji I and Sauer K (1993) Energy transfer dynamics of an isolated light-harvesting complex of Photosystem I from spinach. Time resolved fluorescence measurements at 295 K and 77 K. Biochim Biophys Acta 1142: 311–320

    Article  Google Scholar 

  • Mullet JE, Burke JJ and Arntzen CJ (1980) Chlorophyll proteins of Photosystem I. Plant Physiol 65: 814–822

    Article  PubMed  CAS  Google Scholar 

  • Peters FALJ, van Wielink JE, Wong Fong Sang HW, De Vries S and Kraayenhof R (1983) Studies on well coupled Photosystem I-enriched subchloroplast vesicles. Content and redox properties of electron transfer components. Biochim Biophys Acta 722: 460–470

    Article  CAS  Google Scholar 

  • Palsson LO, Tjus SE, Anderson B and Gillbro T (1995) Ultrafast energy transfer dynamics resolved in isolated spinach light-harvesting complex I and LHC I-730 subpopulation. Biochim Biophys Acta 1230: 1–9

    Article  Google Scholar 

  • Pålsson LO, Fleming F, Gobets BH, van Grondelle R, Dekker JP and Schlodder E (1998) Energy transfer and charge separation in Photosystem I: P700 oxidation upon selective excitation of the long-wavelength antenna chlorophylls of Synechococcus elongatus. Biophys J 74: 2611–2622

    PubMed  Google Scholar 

  • Tapie P, Choquet Y, Breton J, Delepelaire P and Wollman FA (1984) Orientation of Photosystem I pigments. Investigation by low-temperature linear dichroism and polarized fluorescence emission. Biochim Biophys Acta 767: 57–69

    Article  CAS  Google Scholar 

  • Trinkunas G and Holzwarth AR (1994) Kinetic modeling of exciton migration in photosynthetic systems. 2: Simulations of excitation dynamics in two-dimensional Photosystem I core antenna reaction center complexes. Biophys J 66: 415–429

    PubMed  CAS  Google Scholar 

  • van der Lee J, Bald D, Kwa SLS, van Grondelle R, Rogner M and Dekker JP (1993) Steady-state polarized light spectroscopy of isolated Photosystem I complexes. Photosynth Res 35: 311–321

    Article  CAS  Google Scholar 

  • van Grondelle R, Dekker JP, Gilibro T and Sundstrom V (1994) Energy transfer and trapping in photosynthesis. Biochim Biophys Acta 1187: 1–65

    Article  Google Scholar 

  • van Gurp MG, van Ginkel and Levine YK (1989) Fluorescence anisotropy of chlorophyll a and chlorophyll b in castor oil. Biochim Biophys Acta 973: 405–413

    CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Faculty of Physics, Department of Condensed Matter of Physics, Sofia University, 5, J. Bourchier blvd, BG-1164, Sofia, Bulgaria

    Atanaska Andreeva

  2. lnstitute of Biophysics, Bulgarian Academy of Sciences, Acad G. Bonchev str. bl.21, 1113, Sofia, Bulgaria

    Maya Velitchkova

Authors
  1. Atanaska Andreeva
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Maya Velitchkova
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Atanaska Andreeva.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Andreeva, A., Velitchkova, M. Polarized fluorescence spectroscopy of oriented isolated spinach Photosystem I particles. Photosynthesis Research 65, 15–28 (2000). https://doi.org/10.1023/A:1006480915298

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1006480915298

Search

Navigation