Abstract
The protocols regarding separation and purification of LHCII subcomplexes by non-denaturing isoelectric focusing (ndIEF), confirmation of trimeric nature of the subcomplexes, and establishing their polypeptide composition are described below.
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References
Jansson, S. (1999) A guide to the identification of the Lhc genes and their relatives in Arabidopsis. Trends Plant Sci. 4, 236–240.
Butler, J. P. G. and Kuhlbrandt, W. (1988) Determination of the aggregate size in detergent solution of the light-harvesting chlorophyll a/b-protein complex from chloroplast membranes. Proc. Natl. Acad. Sci. USA 85, 3797–3801.
Kuhlbrandt, W. (1987) Three-dimensional crystals of the light-harvesting chlorophyll a/b-protein complex from pea chloroplasts. J. Mol. Biol. 194, 757–762.
Jackowski, G. and Jansson, S. (1998) Characterization of photosystem II antenna complexes separated by non-denaturing isoelectric focusing. Z. Natufrorforsch. 53c, 841–848.
Jackowski, G., Kacprzak, K., and Jansson, S. (2001) Identification of Lhcb1/Lhcb2/Lhcb3 heterotrimers of the main light-harvesting chlorophyll a/b-protein complex of photosystem II (LHCII). Biochim. Biophys. Acta 1504, 340–345.
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.
Dunahay, T. G., Staehelin, L. A., Seibert, M., Ogilvie, P. D., and Berg, S. P. (1984) Structural, biochemical and biophysical characterization of four oxygenevolving photosystem II preparations from spinach. Biochim. Biophys. Acta 764, 179–193.
Enami, I., Kamino, K., Shen, J.-R., Satoh, K., and Katoh, S. (1989) Isolation and characterization of photosystem II complexes which lack light-harvesting chlorophyll a/b proteins but retain three extrinsic proteins related to oxygen evolution from spinach. Biochim. Biophys. Acta 977, 33–39.
Jackowski, G. and Kluck, E. (1994) The oligomeric arrangement of the light-harvesting chlorophyll a/b-protein complex of photosystem II. Z. Naturforsch. 49c, 337–342.
Barber, J. (1980) An explanation for the relationship between salt-induced thylakoid stacking and the chlorophyll flurescence changes associated with changes in spillover of energy from photosystem II to photosystem I. FEBS Lett. 118, 1–10.
Sculley, M. J., Duniec, J. T., Thorne, S. W., Chois, W. S., and Boardman, H. K. (1980) The stacking of chloroplast thylakoids. Quantitative analysis of them balance of ofrces between thylakoid membranes of chloroplasts and the role of divalent cations. Arch. Biochem. Biophys. 201, 339–346.
Jackowski, G. and Przymusinski, R. (1995). The resolution and biochemical characterization of subcomplexes of the main light-harvesting chlorophyll a/b-protein complex of photosystem II (LHCII). Photosynth. Res. 43, 41–48.
Jackowski, G. (1996) The subcomplex organization of the major chlorohyll a/b-protein light-harvesting complex of photosystem II (LHCII) in barley thylakoid membrane. Z. Naturforsch. 51c, 454–463.
Hobe, S., Prytulla, S., Kuhlbrandt, W., and Paulsen, H. (1994) Trimerization and crystallization of reconsituted light-harvesting chlorophyll a/b complex. EMBO J. 13, 3423–3429.
Nussberger, S., Dekker, J. P., Kuhlbrandt, W., Van Bolhuis, B. M., Van Grondelle, R., and Van Amerongen, H. (1994) Spectroscopic characterization of three different monomeric forms of the main chlorohyll a/b binding protein from chloroplast membranes. Biochemistry 33, 14,775–14,783.
Laemmli, U. K. (1970) Cleavege of structural proteins during the assembly of the head of bacteriophage T4. Nature 277, 680–685.
O’Farell, P. H. (1975) High resolution two-dimensional electrophoresis of proteins. J. Biol. Chem. 250, 4007–4021.
Jansson, S., Stefansson, H., Nystrom, U., Gustaffosn, P. A., and Albertsson, P.-A. (1997) Antenna protein composition of PSI and PSII in thylakoid sub-domains. Biochim. Biophys. Acta 1320, 297–309.
Neuhoff, V., Arold, N., Taube, D., and Erhardt, W. (1988) Improved staining of proteins in polyacrylamide gels including isoelectric focusing gels with Coomassie Brilliant Blue G-250 and R-250. Electrophoresis 9, 255–262.
Blakesley, R. W. and Boezi, J. A. (1977) A new staining technique for proteins in polyacrylamide gels using Coomassie Brilliant Blue G-250. Anal. Biochem. 82, 580–582.
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Jackowski, G. (2004). Separation, Purification, and Characterization of Polypeptide Composition of Subcomplexes of the Main Light-Harvesting Chlorophyll a/b-Protein Complex of Photosystem II. In: Carpentier, R. (eds) Photosynthesis Research Protocols. Methods In Molecular Biology™, vol 274. Humana Press. https://doi.org/10.1385/1-59259-799-8:115
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DOI: https://doi.org/10.1385/1-59259-799-8:115
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