Skip to main content
SpringerLink
Log in
Book cover

Photosynthesis Research Protocols pp 183–204Cite as

Photosystem II Reconstitution Into Proteoliposomes

Structure-Function Characterization

  • Protocol

Part of the book series: Methods In Molecular Biology™ ((MIMB,volume 274))

Abstract

This chapter discusses the photosystem II (PSII) reconstitution into proteoliposomes. In the first part of the chapter, protocols are outlined (1) for the preparation of lipid bilayer vesicles (liposomes) constituted of individual thylakoid lipids or their mixtures (2) for the preparation of PSII particles, and (3) for the incorporation of the PSII particles into the liposomes. In the second part of the chapter, methodologies are described for the structure-function characterization of the PSII-lipid complexes (proteoliposomes). This includes the sodium dodecyl sulfate-poly-acrylamide gel electrophoresis (SDS-PAGE) determination of the PSII proteins, measurement of oxygen-evolving activity of PSII in the proteoliposomes, and the study of structural changes of the PSII proteins upon their incorporation into the lipid bilayers by Fourier transform infrared (FTIR) spectroscopy.

This is a preview of subscription content, log in via an institution.

Protocol
USD   49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   74.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever

Tax calculation will be finalised at checkout

Purchases are for personal use only

Learn about institutional subscriptions

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Joyard, J., Maréchal, E., Miège, C., Block, M. A., Dorne, A.-J., and Douce, R. (1998) Structure, distribution and biosynthesis of glycerolipids from higher plant chloroplasts, in Lipids in Photosynthesis: Structure, Function and Genetics (Siegenthaler, P. A. and Murata, N., eds.), Kluwer Academic Publishers, Dordrecht, pp. 21–52.

    Google Scholar 

  2. Williams, W. P. (1998) The physical properties of thylakoid membrane lipids and their relation to photosysnthesis, in Lipids in Photosynthesis: Structure, Function and Genetics (Siegenthaler, P. A. and Murata, N., eds.), Kluwer Academic Publishers, Dordrecht, pp. 103–118.

    Google Scholar 

  3. Dorne, A.-J., Block, M. A., Joyard, J., and Douce, R. (1990) Do thylakoids really contain phosphatidylcholine? Proc. Natl. Acad. Sci. USA 87, 71–74.

    Article  PubMed  CAS  Google Scholar 

  4. Nénonéné, E. K. and Fragata, M. (1998) Interaction of photosystem II proteins with non-aggregated membranes constituted of phosphatidylglycerol and the electrically neutral phosphatidylcholine enhances the oxygen-evolving activity. Chem. Phys. Lipids 91, 97–107.

    Article  Google Scholar 

  5. Block, M. A., Dorne, A. J., Joyard, J., and Douce, R. (1983) Preparation and characterization of membrane fractions enriched in outer and inner envelope membranes from spinach chloroplasts. II-Biochemical characterization. J. Biol. Chem. 258, 13,281–13,286.

    PubMed  CAS  Google Scholar 

  6. Siegenthaler, P. A., Rawyler, A., and Giroud, C. (1987) Spatial organization and functional roles of acyl lipids in thylakoid membranes, in The Metabolism, Structure and Function of Plant Lipids (Stumpf, P. K., Mudd, J. B., and Nes, W. D., eds.), Plenum, New York and London, pp. 161–168.

    Google Scholar 

  7. Keegstra, K. and Froehlich, J. (1999) Protein import into chloroplasts. Curr. Opin. Plant Biol. 2, 471–476.

    Article  PubMed  CAS  Google Scholar 

  8. Hankamer, B., Barber, J., and Boekema, E. J. (1997) Structure and membrane organization of photosystem II in green plants. Annu. Rev. Plant Physiol. Plant Mol. Biol. 48, 641–671.

    Article  PubMed  CAS  Google Scholar 

  9. Siegenthaler, P. A. and Murata, N. (eds.) (1998) Lipids in Photosynthesis: Structure, Function and Genetics. Kluwer Academic Publishers, Dordrecht.

    Google Scholar 

  10. CSIRO (ed.) (2001) PS2001 Proceedings: 12th International Congress on Photosynthesis. CSIRO Publishing, Melbourne.

    Google Scholar 

  11. Lessard, J. G. and Fragata, M. (1986) Micropolarities of lipid bilayers and micelles. 3. Effect of monovalent ions on the dielectric constant of the water-membrane interface of unilamellar phosphatidylcholine vesicles. J. Phys. Chem. 90, 811–817.

    Article  CAS  Google Scholar 

  12. Fragata, M., Menikh, A., and Robert, S. (1993) Salt-mediated effects in nonionic lipid bilayers constituted of digalactosyldiacylglycerol studied by FTIR spectroscopy and molecular modellization. J. Phys. Chem. 97, 13,920–13,926.

    Article  CAS  Google Scholar 

  13. Menikh, A. and Fragata, M. (1993) Fourier transform infrared spectroscopic study of ion binding and intramolecular interactions in the polar head of digalactosyl diacylglycerol. Eur. Biophys. J. 22, 249–258.

    Article  PubMed  CAS  Google Scholar 

  14. Tanford, C. (1973) The Hydrophobic Effect: Formation of Micelles and Biological Membranes. John Wiley, New York.

    Google Scholar 

  15. Huang, C.-H. (1969) Studies on phosphatidylcholine vesicles. Formation and physical characteristics. Biochemistry 8, 344–351.

    Article  PubMed  CAS  Google Scholar 

  16. Fragata, M., El-Kindi, M., and Bellemare, F. (1985) Mixing of single-chain amphiphiles in two-chain lipid bilayers. Characteristics of chlorophyll a and α-tocopherol incorporation in unilamellar phosphatidylcholine vesicles. Chem. Phys. Lipids 37, 117–125.

    Article  PubMed  CAS  Google Scholar 

  17. Kepczynski, M., Ramasamy, P. P., Smith, K. M., and Ehrenberg, B. (2002) Do liposome-binding constants of porphyrins correlate with their measured and predicted partitioning between octanol and water? Photochem. Photobiol. 76, 127–134.

    Article  PubMed  CAS  Google Scholar 

  18. Zhao, H., Mattila, J.-P., Holopainen, J. M., and Kinnunen, P. K. J. (2001) Comparison of the membrane association of two antimicrobial peptides, magainin 2 and indolicidin. Biophys. J. 81, 2979–2991.

    Article  PubMed  CAS  Google Scholar 

  19. Marsh, D. (1990) CRC Handbook of Lipid Bilayers. CRC Press, Boca Raton.

    Google Scholar 

  20. Garab, G., Lohner, K., Laggner, P., and Farkas, T. (2000) Self-regulation of the lipid content of membranes by non-bilayer lipids: a hypothesis. Trends in Plant Sci. 11, 489–494.

    Article  Google Scholar 

  21. Bellemare, F. and Fragata, M. (1981) Transmembrane distributions of α-tocopherol in single-lamellar mixed lipid vesicles. J. Memb. Biol. 58, 67–74.

    Article  CAS  Google Scholar 

  22. L’Heureux, G. P. and Fragata, M. (1988) Micropolarities of lipid bilayers and micelles. 5. Localization of pyrene in small unilamellar phosphatidylcholine vesicles. Biophys. Chem. 30, 293–301.

    Article  PubMed  Google Scholar 

  23. Berthold, D. A., Babcock, G. T., and Yocum, C. F. (1981) A highly resolved oxygen-evolving photosystem II preparation from spinach thylakoid membranes. FEBS Lett. 134, 231–234.

    Article  CAS  Google Scholar 

  24. Nénonéné, E. K. and Fragata, M. (1990) Effects of pH and freeze-thaw on photosynthetic oxygen evolution of photosystem II particles incorporated into phosphatidylglycerol bilayers. J. Plant Physiol. 136, 615–620.

    Google Scholar 

  25. Fragata, M., Strzalka, K., and Nénonéné, E. K. (1991) MgCl2-induced reversal of oxygen evolution decay in photosystem II particles incubated with phosphatidylglycerol vesicles at high lipid/photosystem II ratio. J. Photochem. Photobiol. B: Biol. 11, 329–342.

    Article  CAS  Google Scholar 

  26. Arnon, D. I. (1949) Copper enzymes in isolated chloroplasts: polyphenyloxidase in Beta vulgaris. Plant Physiol. 14, 1–15.

    Article  Google Scholar 

  27. Ségui, J. A., Maire, V., Gabashvili, I. S., and Fragata, M. (2000) Oxygen evolution loss and structural transitions in photosystem II induced by low intensity UV-B radiation of 280 nm wavelength. J. Photochem. Photobiol. B: Biol. 56, 39–47.

    Article  Google Scholar 

  28. Joshi, M. and Fragata, M. (1999) Heat-induced changes in the photochemical centres and the protein secondary structures of photosystem II studied by variable fluorescence and difference FT-IR spectroscopy. Z. Naturforsch. 54c, 35–43.

    Google Scholar 

  29. Chua, N.-H. (1980) Electrophoretic analysis of chloroplast proteins. Methods in Enzymology 69, 434–446.

    Article  CAS  Google Scholar 

  30. Chitnis, P. R. (2001) Photosystem I: function and physiology. Annu. Rev. Plant Physiol. Plant Mol. Biol. 52, 593–626.

    Article  PubMed  CAS  Google Scholar 

  31. Arrondo, J. L. R., Muga, A., Castresana, J., and Goñi, F. M. (1993) Quantitative studies of the structure of proteins in solution by Fourier-transform infrared spectroscopy. Prog. Biophys. Mol. Biol. 59, 23–56.

    Article  PubMed  CAS  Google Scholar 

  32. He, W.-Z., Newell, W. R., Haris, P. I., Chapman, D., and Barber, J. (1991) Protein secondary structure of the isolated photosystem II reaction centre and conformational changes studied by Fourier transform infrared spectroscopy. Biochemistry 30, 4552–4559.

    Article  PubMed  CAS  Google Scholar 

  33. MacDonald, G. M. and Barry, B. A. (1992) Difference FT-IR study of a novel biochemical preparation of photosystem II. Biochemistry 31, 9848–9856.

    Article  PubMed  CAS  Google Scholar 

  34. De Las Rivas, J. and Barber, J. (1997) Structure and thermal stability of photosystem II reaction centres studied by infrared spectroscopy. Biochemistry 36, 8897–8903.

    Article  PubMed  Google Scholar 

  35. Gabashvili, I. S., Menikh, A., Ségui, J., and Fragata, M. (1998) Protein structure of photosystem II studied by FT-IR spectroscopy. Effect of digalactosyldiacylglycerol on the tyrosine side chain residues. J. Mol. Struct. 444, 123–133.

    Article  CAS  Google Scholar 

  36. Arrondo, J. L. R., Castresana, J., Valpuesta, J. M., and Goõn, F. M. (1994) Structure and thermal denaturation of crystalline and mon crystalline cytochrome oxidase as studied by infrared spectroscopy. Biochemistry 33, 11,650–11,655.

    Google Scholar 

  37. Krimm, S. and Bandekar, J. (1986) Vibrational spectroscopy and conformation of peptides, polypeptides, and proteins. Adv. Protein Chem. 38, 181–364.

    Article  PubMed  CAS  Google Scholar 

  38. Surewicz, W. K. and Mantsch, H. H. (1988) New insight into protein secondary structure from resolution-enhanced infrared spectra. Biochim. Biophys. Acta 952, 115–130.

    Article  PubMed  CAS  Google Scholar 

  39. Wilson, E. B. Jr., Decius, J. C., and Cross, P. C. (1955) The Theory of Infrared and Raman Vibrational Spectra. McGraw-Hill, New York.

    Google Scholar 

  40. Menikh, A. and Fragata, M. (1994) Protein secondary structure of the photosystem II complex studied by FT-IR spectroscopy. Changes in the amide I region mediated by digalactosyl diacylglycerol and divalent cations. J. Mol. Struct. 319, 101–107.

    Article  CAS  Google Scholar 

  41. Fragata, M., Nénonéné, E. K., Maire, V., and Gabashvili, I. S. (1997) Structure of the phosphatidylglycerol-photosystem II complex studied by FT-IR spectroscopy. Mg(II) effect on the polar head group of phosphatidylglycerol. J. Mol. Struct. 405, 151–158.

    Article  CAS  Google Scholar 

  42. Heimburg, T. and Marsh, D. (1993) Investigation of secondary and tertiary structural changes of cytochrome c with anionic lipids using amide hydrogen exchange measurements: A FTIR study. Biophys. J. 65, 2408–2417.

    Article  PubMed  CAS  Google Scholar 

  43. Lee, D. C., Haris, P. I., Chapman, D., and Mitchell, R. C. (1990) Determination of protein secondary structure using factor analysis of infrared spectra. Biochemistry 29, 9185–9193.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Département de Chimie-Biologie (GREIB), Université du Québec à Trois-Rivières, Québec, Canada

    Mário Fragata

Authors
  1. Mário Fragata
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. Départment de Chimie-Biologie (GREIB), Université du Québec à Trois-Rivières, Trois-Rivières, Québec, Canada

    Robert Carpentier

Rights and permissions

Reprints and permissions

Copyright information

© 2004 Humana Press Inc., Totowa, NJ

About this protocol

Cite this protocol

Fragata, M. (2004). Photosystem II Reconstitution Into Proteoliposomes. In: Carpentier, R. (eds) Photosynthesis Research Protocols. Methods In Molecular Biology™, vol 274. Humana Press. https://doi.org/10.1385/1-59259-799-8:183

Download citation

  • DOI: https://doi.org/10.1385/1-59259-799-8:183

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-232-2

  • Online ISBN: 978-1-59259-799-4

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation