Comparison of the 伪 and 尾 isomeric forms of the detergent n-dodecyl-D-maltoside for solubilizing photosynthetic complexes from pea thylakoid membranes

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• 作者：Cristina Pagliano^a ; ^{cristina.pagliano@polito.it} ; Simone Barera^a ; Fabiana Chimirri^a ; Guido Saracco^a ; James Barber^a ; ^b ; ^{j.barber@imperial.ac.uk}
• 关键词：ATP-ase ; Adenosine triphosphate synthase ; BN-PAGE ; Blue native polyacrylamide gel electrophoresis ; Chl ; chlorophyll ; CMC ; critical micelle concentration ; Cyt ; cytochrome ; DGDG ; digalactosyldiacylglycerol ; 伪-DM ; n-dodecyl-伪-D-maltoside ; 尾-DM ; n-dodecyl-
• 刊名：Biochimica et Biophysica Acta (BBA)/Bioenergetics
• 出版年：2012
• 期刊代码：13_00052728
• 类别：bio
• 出版时间：August, 2012
• 卷：1817
• 期：8
• 页码：1506-1515
• 文件大小：1192 K

摘要

Mild non-ionic detergents are indispensable in the isolation of intact integral membrane proteins and protein-complexes from biological membranes. Dodecylmaltoside (DM) belongs to this class of detergents being 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 center of the carbohydrate head group, axial in 伪-DM and equatorial in 尾-DM. In this paper, we have investigated the solubilizing properties of 伪-DM and 尾-DM on the isolation of photosynthetic complexes from pea thylakoids membranes maintaining their native architecture of stacked grana and stroma lamellae. Exposure of these stacked thylakoids to a single step treatment with increasing concentrations (5-100 mM) of 伪-DM or 尾-DM resulted in a quick partial or complete solubilization of the membranes. Regardless of the isomeric form used: 1) at the lowest DM concentrations only a partial solubilization of thylakoids was achieved, giving rise to the release of mainly small protein complexes mixed with membrane fragments enriched in PSI from stroma lamellae; 2) at concentrations above 30 mM a complete solubilization occurred with the further release of high molecular weight protein complexes identified as dimeric PSII, PSI-LHCI and PSII-LHCII supercomplexes. However, at concentrations of detergent which fully solubilized the thylakoids, the 伪 and 尾 isomeric forms of DM exerted a somewhat different solubilizing effect on the membranes: higher abundance of larger sized PSII-LHCII supercomplexes retaining a higher proportion of LHCII and lower amounts of PSI-LHCI intermediates were observed in 伪-DM treated membranes, reflecting the mildness of 伪-DM compared with its isomer. This article is part of a Special Issue entitled: Photosynthesis Research for Sustainability: from Natural to Artificial.