用非变性去污剂方法制备高度富集的光系统 II 膜囊泡。
Preparation of highly enriched photosystem II membrane vesicles by a non-detergent method.
机构信息
Department of Biochemistry, University of Lund, P.O. Box 124, S-221 00, Lund, Sweden.
出版信息
Photosynth Res. 1989 Jun;20(3):249-59. doi: 10.1007/BF00034068.
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)).
介绍了一种改良的、非去垢剂的从菠菜叶绿体中制备分离 PS II 膜囊泡的方法。通过 Yeda 压处理和水相两相分配将类囊体（叶绿素 (Chl) a/b 比 2.8，Chl/P700 435）分级，得到外翻囊泡 (1)（chl a/b 2.2，chl/P700 700）。这些囊泡进行超声 - 相分离程序；外翻囊泡的超声处理步骤，同时仍存在于葡聚糖-聚乙二醇两相系统中，通过相分离交替进行。这些步骤从外翻囊泡中选择性地去除了含有 P700 的膜片段，并得到了 PS II 纯度提高的膜级分（Chl a/b 比 1.9，Chl/P700 1500），并保持了氧释放能力（295 μmol O2 mg Chl(-1) h(-1)）。
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