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Trehalose-protected lipid membranes for determining membrane protein structure and insertion

机译:海藻糖保护的脂质膜可确定膜蛋白的结构和插入

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Trehalose preserves lipid bilayers during dehydration and rehydration by replacing water to form hydrogen bonds between its own 011 groups and lipid headgroups. We compare the lipid conformation and dynamics between trehalose-protected 1yophilized membranes and hydrated membranes, to assess the suitability of the trehalose-containing membrane as a matrix for membrane protein structure determination. P-31 spectra indicate that the lipid headgroup of trehalose-protected dry POPC membrane (TRE-POPC) have an effective phase transition temperature that is similar to 50 K higher than that of the hydrated POPC membrane. In contrast, the acyl chains have similar transition temperatures in the two membranes. Intramolecular lipid C-13-P-31 distances are the same in TRE-POPC and crystalline POPC, indicating that the lipid headgroup and glycerol backbone conformation is unaffected by trehalose incorporation. Intermolecular C-13-P-31 distances between a membrane peptide and the lipid headgroups are 10% longer in the hydrated membrane at 226 K than in the trehalose-protected dry membrane at 253 K. This is attributed to residual motions in the hydrated membrane, manifested by the reduced P-31 chemical shift anisotropy, even at the low temperature of 226 K. Thus, trehalose lyoprotection facilitates the study of membrane protein structure by allowing experiments to be conducted at higher temperatures than possible with the hydrated membranes. (c) 2006 Elsevier Inc. All rights reserved.
机译:海藻糖通过替换水以在其自身的011基团和脂质头基之间形成氢键,从而在脱水和再水化过程中保留脂质双层。我们比较了海藻糖保护的1冻干膜和水合膜之间的脂质构象和动力学,以评估含海藻糖膜作为膜蛋白结构测定基质的适用性。 P-31光谱表明,海藻糖保护的干式POPC膜(TRE-POPC)的脂质头基具有比水合POPC膜高50 K的有效相变温度。相反,酰基链在两个膜中具有相似的转变温度。在TRE-POPC和结晶POPC中,分子内脂质C-13-P-31的距离相同,表明脂质头基和甘油主链构象不受海藻糖掺入的影响。膜肽和脂质头基之间的分子间C-13-P-31距离在226 K的水合膜中比在253 K的海藻糖保护的干膜中长10%。这归因于在水合膜中的残留运动,即使在226 K的低温下,P-31的化学位移各向异性也降低了。因此,海藻糖的冻干保护通过允许在比水合膜更高的温度下进行实验,促进了膜蛋白结构的研究。 (c)2006 Elsevier Inc.保留所有权利。

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