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对正常细胞、肿瘤细胞及其相应质膜进行荧光偏振测量。

Fluorescence polarization measurements on normal and tumour cells and their corresponding plasma membranes.

作者信息

van Hoeven R P, van Blitterswijk W J, Emmelot P

出版信息

Biochim Biophys Acta. 1979 Feb 20;551(1):44-54. doi: 10.1016/0005-2736(79)90351-1.

Abstract

Using 1,6-diphenyl-1,3,5-hexatriene as a probe, the degree of fluorescence polarization (P) at 25 degrees C of intact and disrupted cells and isolated plasma membranes were compared for a variety of systems. 1. Human erythrocytes, mouse thymocyte and leukemia cells, rat liver and hepatoma cells, and human and mouse milk fat globules displayed P values ranging from 0.300 to 0.120. 2. P values or probe labelling rates of intact and disrupted cells were similar. 3. As compared with whole or disrupted cells, the higher to much higher P values of plasma membranes isolated from the corresponding cells showed only a limited mutual variation. 4. delta P values, being the difference in P values between plasma membranes and whole cells were attributed to the extent to which endomembranes and non-membrane lipids contributed. Among these, triglycerides had the greatest relative effect. 5. Though a particular isolation procedure for plasma membranes may select for more rigid fragments, this effect is by far not sufficient to account for the observed delta P values. It is concluded that the fluorescence polarization technique with a lipophilic probe applied to whole cells represents a measure of the average fluidity of all lipids being present in a cell and thus does not exclusively monitor the cell surface membrane.

摘要

以1,6 - 二苯基 - 1,3,5 - 己三烯为探针,比较了多种体系中完整细胞、破碎细胞及分离的质膜在25℃时的荧光偏振度(P)。1. 人红细胞、小鼠胸腺细胞和白血病细胞、大鼠肝细胞和肝癌细胞以及人和小鼠乳脂肪球的P值范围为0.300至0.120。2. 完整细胞和破碎细胞的P值或探针标记率相似。3. 与完整或破碎细胞相比,从相应细胞分离的质膜的P值较高至高得多,仅显示出有限的相互变化。4. ΔP值(即质膜和完整细胞之间P值的差异)归因于内膜和非膜脂质的贡献程度。其中,甘油三酯的相对影响最大。5. 尽管质膜的特定分离程序可能会选择更刚性的片段,但这种影响远不足以解释观察到的ΔP值。得出结论,将亲脂性探针应用于完整细胞的荧光偏振技术代表了对细胞中所有脂质平均流动性的一种测量,因此并非专门监测细胞表面膜。

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