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Rapid and simple isolation of pure photosystem II core and reaction center particles from spinach
Authors:Peter J. van Leeuwen  Maaike C. Nieveen  Erik Jan van de Meent  Jan P. Dekker  Hans J. van Gorkom
Affiliation:(1) Department of Biophysics, Huygens laboratory of the State University, P.O. Box 9504, 2300 RA Leiden;(2) Department of Physics and Astronomy, Free University, De Boelelaan 1081, 1081 HV Amsterdam, the Netherlands
Abstract:Pure and active oxygen-evolving PS II core particles containing 35 Chl per reaction center were isolated with 75% yield from spinach PS II membrane fragments by incubation with n-dodecyl-beta-D-maltoside and a rapid one step anion-exchange separation. By Triton X-100 treatment on the column these particles could be converted with 55% yield to pure and active PS II reaction center particles, which contained 6 Chl per reaction center.Abbreviations Bis-Tris bis[2-hydroxyethyl]imino-tris[hydroxymethyl]methane - Chl chlorophyll - CP29 Chl a/b protein of 29 kDa - Cyt b559 cytochrome b559 - DCBQ 2,5-dichloro-p-benzo-quinone - LHC II light-harvesting complex II, predominant Chl a/b protein - MES 2-[N-Morpholino]ethanesulfonic acid - Pheo pheophytin - PS H photosystem II - QA bound plastoquinone, serving as the secondary electron acceptor in PS II (after Pheo) - SDS sodiumdodecylsulfate
Keywords:photosystem II  core  reaction center  isolation
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