荧光偏振分辨成像探测 MHC Ⅰ类蛋白和脂探针在细胞膜中的取向行为。
Probing orientational behavior of MHC class I protein and lipid probes in cell membranes by fluorescence polarization-resolved imaging.
机构信息
Aix-Marseille Université, Ecole Centrale Marseille, Centre National de la Recherche Scientifique, Institut Fresnel, Marseille, France.
出版信息
Biophys J. 2011 Jul 20;101(2):468-76. doi: 10.1016/j.bpj.2011.05.021.
Abstract
Steady-state polarization-resolved fluorescence imaging is used to analyze the molecular orientational order behavior of rigidly labeled major histocompatibility complex class I (MHC I) proteins and lipid probes in cell membranes of living cells. These fluorescent probes report the orientational properties of proteins and their surrounding lipid environment. We present a statistical study of the molecular orientational order, modeled as the width of the angular distribution of the molecules, for the proteins in the cell endomembrane and plasma membrane, as well as for the lipid probes in the plasma membrane. We apply this methodology on cells after treatments affecting the actin and microtubule networks. We find in particular opposite orientational order changes of proteins and lipid probes in the plasma membrane as a response to the cytoskeleton disruption. This suggests that MHC I orientational order is governed by its interaction with the cytoskeleton, whereas the plasma membrane lipid order is governed by the local cell membrane morphology.
摘要
稳态偏振分辨荧光成像用于分析活细胞细胞膜中刚性标记的主要组织相容性复合体 I 类 (MHC I) 蛋白和脂质探针的分子取向有序行为。这些荧光探针报告蛋白质及其周围脂质环境的取向特性。我们对细胞内膜和质膜中蛋白质以及质膜中脂质探针的分子取向有序性进行了统计研究，将其建模为分子角分布的宽度。我们将这种方法应用于影响肌动蛋白和微管网络的细胞处理后。我们发现，特别是作为细胞骨架破坏的响应，质膜中的蛋白质和脂质探针的取向有序性发生了相反的变化。这表明 MHC I 的取向有序性受其与细胞骨架的相互作用控制，而质膜脂质的有序性受局部细胞膜形态的控制。
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