Surface electrostatics of lipid bilayers by EPR of a pH-sensitive spin-labeled lipid.

Surface electrostatics of lipid bilayers by EPR of a pH-sensitive spin-labeled lipid.
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通过 pH 敏感的自旋标记脂质的 EPR 分析脂质双层的表面静电。

DOI:
10.1016/j.bpj.2012.11.3806
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发表时间:
2013
影响因子:
3.4
通讯作者:
Smirnov,AlexI
Smirnov,AlexI
中科院分区:
生物学3区
文献类型:
--
作者:
Voinov,MaximA;Rivera-Rivera,Izarys;Smirnov,AlexI

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许多生物物理过程,如蛋白质插入膜和膜融合,都是由双分子静电势控制的。在撰写本文时,用于这类测量的生物物理方法仅限于几种技术。在这里,据我们所知,我们描述了一种新的自旋探针电子顺磁共振(EPR)方法来评估脂质双层的静电表面电位,该方法基于最近合成的EPR探针(IMTSL-PTE),该探针包含一个可可逆电离的氮氧化物标记,该标记连接在脂类的极性头基上。探针的EPR谱直接报告其电离状态,从而通过改变氮氧化物的磁性参数和旋转平均程度来报告静电势。此外,探针的脂类性质提供了它与脂双层的完全整合。将氮氧化物部分直接系接到脂极性头基上定义了测量的电势相对于脂双层界面的位置。IMTSL-PTE的EPR测量表明,在低电解质浓度(50 Mm)下,DMPG阴离子双层膜在液体相(48°C)和凝胶相(17℃)中的静电表面电位与Gouy-Chapman理论的一致性在±2%以内。对于凝胶相(17°C)的DMPG囊泡,随着DMPG/DMPC和POPG/POPC混合物形成的电解质浓度和流体相双层的不同,这种一致性开始减弱。讨论了产生这种偏差的可能原因,以及静电中性参考界面的适当选择。所描述的EPR方法有望完全适用于更复杂的细胞膜模型。
Many biophysical processes such as insertion of proteins into membranes and membrane fusion are governed by bilayer electrostatic potential. At the time of this writing, the arsenal of biophysical methods for such measurements is limited to a few techniques. Here we describe a, to our knowledge, new spin-probe electron paramagnetic resonance (EPR) approach for assessing the electrostatic surface potential of lipid bilayers that is based on a recently synthesized EPR probe (IMTSL-PTE) containing a reversibly ionizable nitroxide tag attached to the lipids' polar headgroup. EPR spectra of the probe directly report on its ionization state and, therefore, on electrostatic potential through changes in nitroxide magnetic parameters and the degree of rotational averaging. Further, the lipid nature of the probe provides its full integration into lipid bilayers. Tethering the nitroxide moiety directly to the lipid polar headgroup defines the location of the measured potential with respect to the lipid bilayer interface. Electrostatic surface potentials measured by EPR of IMTSL-PTE show a remarkable (within ±2%) agreement with the Gouy-Chapman theory for anionic DMPG bilayers in fluid (48°C) phase at low electrolyte concentration (50 mM) and in gel (17°C) phase at 150-mM electrolyte concentration. This agreement begins to diminish for DMPG vesicles in gel phase (17°C) upon varying electrolyte concentration and fluid phase bilayers formed from DMPG/DMPC and POPG/POPC mixtures. Possible reasons for such deviations, as well as the proper choice of an electrostatically neutral reference interface, have been discussed. Described EPR method is expected to be fully applicable to more-complex models of cellular membranes.
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发表时间: 2002
期刊: The journal of physical chemistry. B
影响因子: --
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发表时间: 1989-02-13
期刊: BIOCHIMICA ET BIOPHYSICA ACTA
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DOI: 10.1006/jmrb.1996.0073
发表时间: 1996
期刊: Journal of magnetic resonance. Series B
影响因子: --
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