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两相脂质双层中渗滤和区室化的荧光猝灭研究。

Fluorescence-quenching study of percolation and compartmentalization in two-phase lipid bilayers.

作者信息

Piknová B, Marsh D, Thompson T E

机构信息

Department of Biochemistry, University of Virginia, Charlottesville 22908, USA.

出版信息

Biophys J. 1996 Aug;71(2):892-7. doi: 10.1016/S0006-3495(96)79291-3.

Abstract

Fluorescence quenching of a lipid-labeled fluorophore by a lipid spin-labeled quencher has been studied experimentally in two-component, two-phase phosphatidylcholine bilayers to examine the effect of phase connection and disconnection on quenching. Both fluorophore and quencher prefer the fluid phase. At the percolation threshold, the point at which the fluid phase becomes subdivided into may small disconnected domains, the quenching drops abruptly. This decrease in quenching is a function of the fluid-phase fraction and is due to the heterogeneous distribution of fluorophores and quenchers over the fluid-phase domains. Computer simulations of the system were carried out with a triangular lattice divided into closed compartments of variable size and reactant occupancy. The simulations demonstrate that the degree of quenching is reduced in the disconnected systems and that the reduction is correlated with the size of the disconnected domains. The combination of experimental data with simulations leads to the conclusion that at constant temperature the size of fluid-phase domains, nfluid, in the region of the coexistence of the fluid and gel phases is proportional to the fluid fraction, Xfluid. This is in a qualitative agreement with a previous electron spin resonance study of interlipid spin-spin interactions in the same two-component, two-phase bilayer system.

摘要

通过脂质自旋标记猝灭剂对脂质标记荧光团的荧光猝灭已在双组分、两相磷脂酰胆碱双层膜中进行了实验研究,以考察相连接和相断开对猝灭的影响。荧光团和猝灭剂都倾向于流体相。在逾渗阈值处,即流体相开始细分为许多小的不相连区域的点,猝灭突然下降。猝灭的这种降低是流体相分数的函数,并且是由于荧光团和猝灭剂在流体相区域上的不均匀分布所致。使用分为大小可变和反应物占据情况的封闭隔室的三角晶格对该系统进行了计算机模拟。模拟结果表明,在不相连的系统中猝灭程度降低,并且这种降低与不相连区域的大小相关。实验数据与模拟结果相结合得出结论:在恒定温度下,流体相和凝胶相共存区域中流体相区域的大小(n_{fluid})与流体分数(X_{fluid})成正比。这与先前对同一双组分、两相双层膜系统中脂质间自旋 - 自旋相互作用的电子自旋共振研究在定性上是一致的。

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