Abstract
This chapter contains the description of several methods used for the isolation of functional photosystem (PS)II core particles from wild-type (wt), PSI-less, and CP47 histidine-tagged cells of the cyanobacterium Synechocystis sp. PCC 6803. These protocols discuss the cultivation of PSI-containing and PSI-less cells, isolation of thylakoid membranes, purification of PSII core particles using a weak cation exchange or metal affinity column chromatography, and characterization of the final preparation. The described isolation procedures, which normally yield PSII particles highly active in oxygen evolution, can be easily adapted for obtaining preparations from different types of Synechocystis mutants with modified PSII.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Rhee, K. H. (2001) Photosystem II: the solid structural era. Annu. Rev. Biophys. Biomol. Struct. 30, 307–328.
Diner, B. A. and Rappaport, F. (2002) Structure, dynamics, and energetics of the primary photochemistry of photosystem II of oxygenic photosynthesis. Annu. Rev. Plant Biol. 53, 551–580.
Vermaas, W. F. J. (1998) Gene modifications and mutation mapping to study the function of photosystem II. Method. Enzymol. 297, 293–310.
Vermaas, W. (1996) Molecular genetics of the cyanobacterium Synechocystis sp. PCC 6803: principles and possible biotechnology applications. J. Appl. Phycol. 8, 263–273.
Tang, X. S. and Diner, B. A. (1994) Biochemical and spectroscopic characterization of a new oxygen-evolving photosystem II core complex from the cyanobacterium Synechocystis PCC 6803. Biochemistry 33, 4594–4603.
Bricker, T. M., Morvant, J., Masri, N., Sutton, H. M., and Frankel, L. K. (1998) Isolation of a highly active Photosystem II preparation from Synechocystis 6803 using a histidine-tagged mutant of CP 47. Biochim. Biophys. Acta 1409, 50–57.
Vavilin, D., Xu, H., Lin, S., and Vermaas, W. (2003) Energy and electron transfer in photosystem II of a chlorophyll b-containing Synechocystis sp. PCC 6803 mutant. Biochemistry 42, 1731–1746.
Szabo, I., Rigoni, F., Bianchetti, M., et al. (2001) Isolation and characterization of photosystem II subcomplexes from cyanobacteria lacking photosystem I. Eur. J. Biochem. 268, 5129–5134.
Rögner, M., Nixon, P. J., and Diner, B. A. (1990) Purification and characterization of photosystem I and photosystem II core complexes from wt and phycocyanindeficient strains of the cyanobacterium Synechocystis PCC 6803. J. Biol. Chem. 265, 6189–6196.
Xu, H., Vavilin, D., and Vermaas, W. (2001) Chlorophyll b can serve as the major pigment in functional photosystem II complexes of cyanobacteria. Proc. Natl. Acad. Sci. USA 98, 14,168–14,173.
Shen, G.-Z., Boussiba, S., and Vermaas, W. F. J. (1993) Synechocystis sp. PCC 6803 strains lacking photosystem I and phycobilisome function. Plant Cell 5, 1853–1863.
Rippka, R., Deruelles, J., Waterbury, J. B., Herdmann, M., and Stanier, R. Y. (1979) Generic assignments, strain histories and properties of pure cultures of cyanobacteria. J. Gen. Microbiol. 111, 1–61.
Giorgi, L. B., Nixon, P. J., Merry, S. A. P., et al. (1996) Comparison of primary charge separation in the photosystem II reaction center complex isolated from wt and D1-130 mutants of the cyanobacterium Synechocystis PCC 6803. J. Biol. Chem. 271, 2093–2101.
Kzuk, J., Strzclka, K., and Leblanc, R. M. (1992) Monolayer study of plastoquinones α-tocopherol quinone, their hydroquinone forms and their interaction with monogalactosyldiacylglycerol change-transfer complexes in a mixed monolayer. Biochim. Biophys. Acta 1112, 19–26.
Hiyama, T. and Ke, B. (1972) Difference spectra and extinction coefficients of P700. Biochim. Biophys Acta 267, 160–171.
Jordan, P., Fromme, P., Witt, H. T., Klukas, O., Saenger, W., and Krauss, N. (2001) Three-dimensional structure of cyanobacterial photosystem I at 2.5 A resolution. Nature 411, 909–917.
Porra, R. J. (2002) The chequered history of the development and use of simultaneous equations for the accurate determination of chlorophylls a and b. Photosynth. Res. 73, 149–156.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2004 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Vavilin, D.V. (2004). Isolation of Functional Photosystem II Core Particles From the Cyanobacterium Synechocystis sp. PCC 6803. In: Carpentier, R. (eds) Photosynthesis Research Protocols. Methods In Molecular Biology™, vol 274. Humana Press. https://doi.org/10.1385/1-59259-799-8:037
Download citation
DOI: https://doi.org/10.1385/1-59259-799-8:037
Publisher Name: Humana Press
Print ISBN: 978-1-58829-232-2
Online ISBN: 978-1-59259-799-4
eBook Packages: Springer Protocols