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用α-和β-十二烷基-D-麦芽糖苷一步处理从豌豆类囊体膜中分离 PSII-LHCII 超复合体的特性。

Characterization of PSII-LHCII supercomplexes isolated from pea thylakoid membrane by one-step treatment with α- and β-dodecyl-D-maltoside.

机构信息

Applied Science and Technology Department-BioSolar Laboratory, Politecnico di Torino, Viale T. Michel 5, Alessandria, Italy.

出版信息

Philos Trans R Soc Lond B Biol Sci. 2012 Dec 19;367(1608):3389-99. doi: 10.1098/rstb.2012.0056.

Abstract

It was the work of Jan Anderson, together with Keith Boardman, that showed it was possible to physically separate photosystem I (PSI) from photosystem II (PSII), and it was Jan Anderson who realized the importance of this work in terms of the fluid-mosaic model as applied to the thylakoid membrane. Since then, there has been a steady progress in the development of biochemical procedures to isolate PSII and PSI both for physical and structural studies. Dodecylmaltoside (DM) has emerged as an effective mild detergent for this purpose. DM is a glucoside-based surfactant with a bulky hydrophilic head group composed of two sugar rings and a non-charged alkyl glycoside chain. Two isomers of this molecule exist, differing only in the configuration of the alkyl chain around the anomeric centre of the carbohydrate head group, axial in α-DM and equatorial in β-DM. We have compared the use of α-DM and β-DM for the isolation of supramolecular complexes of PSII by a single-step solubilization of stacked thylakoid membranes isolated from peas. As a result, we have optimized conditions to obtain homogeneous preparations of the C(2)S(2)M(2) and C(2)S(2) supercomplexes following the nomenclature of Dekker & Boekema (2005 Biochim. Biophys. Acta 1706, 12-39). These PSII-LHCII supercomplexes were subjected to biochemical and structural analyses.

摘要

是 Jan Anderson 与 Keith Boardman 共同的工作表明,将光系统 I (PSI) 与光系统 II (PSII) 从物理上分离是可行的,而正是 Jan Anderson 意识到了这项工作对于应用于类囊体膜的流动镶嵌模型的重要性。从那时起,为了进行物理和结构研究,不断发展出用于分离 PSII 和 PSI 的生化程序。十二烷基麦芽糖苷 (DM) 已成为一种有效的温和去污剂。DM 是一种基于葡萄糖的表面活性剂,具有由两个糖环和一个不带电荷的烷基糖苷链组成的庞大亲水头部基团。这种分子存在两种异构体,仅在碳水化合物头部基团的非环烷基糖苷链围绕差向中心的构型上有所不同,α-DM 为轴向,β-DM 为赤道向。我们比较了使用α-DM 和β-DM 通过一步溶解从豌豆中分离的堆叠类囊体膜来分离 PSII 的超分子复合物。结果,我们优化了条件,以获得按照 Dekker 和 Boekema(2005 Biochim. Biophys. Acta 1706, 12-39)命名的 C(2)S(2)M(2) 和 C(2)S(2) 超复合物的均相制剂。这些 PSII-LHCII 超复合物进行了生化和结构分析。

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