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用于研究核孔蛋白膜相互作用的巨型单层囊泡(GUVs)

Giant Unilamellar Vesicles (GUVs) to Study Membrane Interaction of Nucleoporins.

作者信息

Tatarek-Nossol Marianna, Antonin Wolfram

机构信息

Institute of Biochemistry and Molecular Cell Biology, Medical School, RWTH Aachen University, Aachen, Germany.

出版信息

Methods Mol Biol. 2025;2958:239-253. doi: 10.1007/978-1-0716-4714-1_16.

Abstract

Giant unilamellar vesicles (GUVs) are large, spherical lipid bilayer vesicles typically ranging from 1 to 100 micrometers in diameter, far exceeding the size of typical small unilamellar vesicles (SUVs) employed, for example, in liposome floatation assays. Their unique size and structural simplicity make them particularly suitable for a wide range of biophysical studies. GUVs serve as versatile model systems for studying membrane-protein interactions, membrane curvature sensing, and membrane remodeling processes. In addition, by micromanipulation, fluorescence recovery after photobleaching, and single-molecule imaging, membrane bending, deformation, and fusion events can be investigated with high spatial and temporal resolution. Here, we summarize our protocols generating GUVs for studying nucleoporin-membrane interaction including protocols reconstituting transmembrane nucleoporins into GUVs.

摘要

巨型单层囊泡(GUVs)是大型球形脂质双层囊泡,直径通常在1至100微米之间,远远超过例如用于脂质体浮选测定的典型小单层囊泡(SUVs)的大小。它们独特的尺寸和结构简单性使其特别适合广泛的生物物理研究。GUVs作为通用模型系统,用于研究膜 - 蛋白相互作用、膜曲率传感和膜重塑过程。此外,通过显微操作、光漂白后的荧光恢复和单分子成像,可以以高空间和时间分辨率研究膜弯曲、变形和融合事件。在这里,我们总结了我们生成用于研究核孔蛋白 - 膜相互作用的GUVs的方案,包括将跨膜核孔蛋白重组到GUVs中的方案。

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