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通过纳秒荧光技术研究脂质双层膜的动态结构。

Dynamic structure of lipid bilayers studied by nanosecond fluorescence techniques.

作者信息

Kawato S, Kinosita K, Ikegami A

出版信息

Biochemistry. 1977 May 31;16(11):2319-24. doi: 10.1021/bi00630a002.

Abstract

Molecular motions in liposomes of dipalmitoyl-phosphatidylcholine (DPPC) were studied by nanosecond fluorescence techniques. As a fluorescent probe for the hydrocarbon region, 1,6-diphenyl-1,3,5-hexatriene (DPH) was used. Time courses of fluorescence intensity IT(t) and emission anisotropy r(t) of DPH embedded in DPPC liposomes were measured at various temperatures. The value of the fluorescence lifetime tau obtained froma single exponential decay of IT(t) was somewhat higher than that in liquid paraffin below the transition temperature Tt and decreased above Tt. Higher values of tau below Tt indicate the almost complete hydrophobic environment. The decay curves of r(t) were separated into two phases: an initial fast decreasing phase of the order of one nanosecond and a second almost constant phase. This indicates that the orientational motion of DPH in the hydrocarbon region is described by a wobbling diffusion restricted by a certain anisotropic potential. The results were analyzed on the model that the wobbling diffusion is confined in a cone with a uniform diffusion constant. Though temperature dependence of the cone angle was sigmoidal, that of the wobbling diffusion constant was like the exponential function. The change in the cone angle at Tt was sharper than that in the wobbling diffusion constant at Tt. Estimated values of the viscosity in the cone were an order of magnitude smaller than the values of "microviscosity" which were estimated from the steady-state emission anisotropy without considering the restrictions on the rotational motion.

摘要

采用纳秒荧光技术研究了二棕榈酰磷脂酰胆碱(DPPC)脂质体中的分子运动。作为烃区的荧光探针,使用了1,6 - 二苯基 - 1,3,5 - 己三烯(DPH)。在不同温度下测量了嵌入DPPC脂质体中的DPH的荧光强度IT(t)和发射各向异性r(t)的时间进程。从IT(t)的单指数衰减得到的荧光寿命tau值在转变温度Tt以下略高于液体石蜡中的值,在Tt以上则降低。Tt以下较高的tau值表明几乎完全是疏水环境。r(t)的衰减曲线分为两个阶段:一个初始的快速下降阶段,时间尺度为一纳秒左右,以及第二个几乎恒定的阶段。这表明烃区中DPH的取向运动由受某种各向异性势限制的摆动扩散来描述。结果在摆动扩散局限于具有均匀扩散常数的圆锥体的模型上进行了分析。尽管圆锥角的温度依赖性呈S形,但摆动扩散常数的温度依赖性类似指数函数。Tt处圆锥角的变化比Tt处摆动扩散常数的变化更明显。圆锥体内估计的粘度值比从稳态发射各向异性估计的“微粘度”值小一个数量级,且未考虑对旋转运动的限制。

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