Abstract
An improved, non-detergent, method for preparative isolation of PS II membrane vesicles from spinach chloroplasts is presented. Thylakoids (chlorophyll (Chl) a/b ratio 2.8, Chl/P700 435) were fractionated by Yeda press treatment and aqueous two-phase partition to yield inside-out vesicles (1) (chl a/b 2.2, chl/P700 700). These vesicles were subjected a sonication — phase partitioning procedure; steps of sonication of inside-out vesicles, while still present in a dextran-polyethylene glycol two-phase system were alternated by phase partition. These steps selectively removed P700-containing membrane fragments from the inside-out vesicles and yielded a membrane fraction with improved PS II purity (Chl a/b ratio 1.9, Chl/P700 1500) and retained oxygen evolving capacity (295 μmol O2 mg Chl-1 h-1).
Similar content being viewed by others
References
Åkerlund H.E., Andersson B. and Albertsson P.Å. 1976 Isolation of photosystem II enriched membrane vesicles from spinach chloroplasts by phase partition. Biochim Biophys Acta 449: 525–535.
Åkerlund H.E. and Andersson B. 1983 Quantitative separation of spinach thylakoids into photosystem II enriched inside-out and photosystem I enriched right-side-out vesicles. Biochim Biophys Acta 725: 35–40.
Albertsson P.Å. 1985 Subfractionation of inside-out thylakoid vesicles and the localization of cytochrome f. Physiol Veg 23: 731–739.
Albertsson P.Å. 1986 Partition of Cell Particles and Macromolecules, 3rd edition, Wiley, New York.
Albertsson P.Å., Svensson P. and Persson A. 1987 Subfractionation of inside-out thylakoid vesicles. Acta Chem Scand B41: 134–136.
Albertsson P.Å. and Svensson P. 1988 Counter-current distribution of sonicated inside-out thylakoid vesicles. Mol Cell Biochem 81: 155–163.
Anderson J.M. and Malkin R. 1982 An ERR study of the lateral organization of electron carriers in chloroplast thylakoids. FEBS Lett: 148: 293–296.
Anderson J.M. and Melis A. 1983 Localization of different photosystems in separate regions of chloroplast membranes. Proc Natl Acad Sci USA 80: 745–749.
Andersson B. and Anderson J.M. 1980 Lateral heterogeneity in the distribution of chlorophyll-protein complexes of the thylakoid membranes of spinach chloroplasts. Biochim Biophys Acta 593: 427–440.
Andersson B. and Haehnel W. 1982 Location of photosystem I and photosystem II reaction centres in different thylakoid regions of stacked chloroplasts. FEBS Lett 146: 13–17.
Andersson B., Sundby C., Åkerlund H.E. and Albertsson P.Å. 1985 Inside-out thylakoid vesicles. An important tool for the characterization of the photosynthetic membrane. Physiol Plant 65: 322–330.
Arnon D.I. 1949 Copper enzymes in isolated chloroplasts. Polyphenoloxidase in Beta vulgaris. Plant Physiol 24: 1–15.
Atta-Asafo-Adjei E. and Dilley R.A. 1985 Plastocyanin stimulation of whole chain and photosystem I electron transport in inside-out thylakoid vesicles. Arch Biochem Biophys 243: 660–667.
Berthhold 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.
Henry L.E.A. and Lindberg-Møller B. 1981 Polypeptide composition of an oxygen evolving photosystem II vesicle from spinach chloroplasts. Carlsberg Res Commun 46: 227–242.
Jacobi G. and Lehmann H. 1968 Die Fragmentierung isolierter Chloroplasten 1. Die functionelle und strukturelle Beurteilung von Fragmenten Ultraschall-behandelter Chloroplasten. Z Pflanzen physiol 59: 457–476.
Laemmli U.K. 1970 Clevage of Structural Proteins during the Assembly of the Head of Bacteriophage T4. Nature 227: 680–685.
Melis A. and Brown J.S. 1980 Stoichiometri of system I and system II reaction centers and of plastoquinone in different photosynthetic membranes. Proc Natl Acad Sci USA 77: 4712–4716.
Melis A. and Homan P.H. 1978 A selective effect of Mg2+ on the photochemistry at one type of reaction center in photosystem II of Chloroplasts. Arch Biochem Biophys 190: 523–536.
Melis A., Svensson P. and Albertsson P.Å. 1986 The domain organization of the chloroplast thylakoid membrane. Localization of photosystem I and of the cytochrome b 6 -f complex. Biochim Biophys Acta 850: 402–412.
Michel J.M. and Michel Wolwertz M.R. 1968 Fractionation of the photosynthetic apparatus from broken spinach chloroplasts by sucrose density-gradient centrifugation. In: Carnegie Inst Year Book 67, pp 508–516.
Sane P.V., Goodchild D.J. and Park P.F. 1970 Characterization of chloroplast photosystems 1 and 2 separated by a non-detergent method. Biochim Biophys Acta 216: 162–178.
Sundby C., Andersson B. and Albertsson P.Å. 1982 Conversion of everted thylakoids into vesicles of normal sidedness exposing the outer grana partition membrane surface. Biochim Biophys Acta 688: 709–719.
Svensson P. and Albertsson P.Å. 1987 Subfractionation of inside-out thylakoid vesicles — Preparation of pure photosystem II particles without using detergent. Progr Photosynth Res II: 281–284.
Vallon O., Wollman F.A. and Olive J. 1986 Lateral distribution of the main protein complexes of the photosynthetic apparatus in Chlamydomonas reinhardtii and in spinach: an immunocytochemical study using intact thylakoid membranes and a PS II enriched membrane preparation. Photobiochem Photobiophys 12: 203–220.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Svensson, P., Albertsson, PÅ. Preparation of highly enriched photosystem II membrane vesicles by a non-detergent method. Photosynth Res 20, 249–259 (1989). https://doi.org/10.1007/BF00034068
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00034068