Isolation of photosystem II-enriched membranes and the oxygen-evolving complex subunit proteins from higher plants.
We describe methods to isolate highly active oxygen-evolving photosystem II (PSII) membranes and core complexes from higher plants, and to purify subunits of the oxygen-evolving complex (OEC). The membrane samples used as the material for various in vitro studies of PSII are prepared by solubilizing thylakoid membranes with the nonionic detergent Triton X-100, and the core complexes are prepared by further solubilization of the PSII membranes with n-dodecyl-β-D-maltoside (β-DDM). The OEC subunit proteins are dissociated from the PSII-enriched membranes by alkaline or salt treatment, and are then purified by ion-exchange chromatography using an automated high performance liquid chromatography system.
Yamamoto
Yasusi
Y
Graduate School of Natural Science and Technology, Okayama University, Okayama, Japan.
Leng
Jing
J
Shen
Jian-Ren
JR
D002458:Cell Fractionation / Q000379:methods*; D002462:Cell Membrane / Q000201:enzymology*; D005591:Chemical Fractionation / Q000379:methods*; D002734:Chlorophyll / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D010100:Oxygen / Q000378:metabolism*; D018532:Pisum sativum / Q000166:cytology* / Q000201:enzymology / Q000378:metabolism; D045332:Photosystem II Protein Complex / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D021122:Protein Subunits / Q000302:isolation & purification* / Q000378:metabolism; D018724:Spinacia oleracea / Q000166:cytology* / Q000201:enzymology / Q000378:metabolism
D045332:Photosystem II Protein Complex; D021122:Protein Subunits; D002734:Chlorophyll; D010100:Oxygen