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Rupturing Giant Plasma Membrane Vesicles to Form Micron-sized Supported Cell Plasma Membranes with Native Transmembrane Proteins

机译:破裂巨型血浆膜囊泡以形成具有天然跨膜蛋白的微米级支持细胞膜

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摘要

Being able to directly obtain micron-sized cell blebs, giant plasma membrane vesicles (GPMVs), with native membrane proteins and deposit them on a planar support to form supported plasma membranes could allow the membrane proteins to be studied by various surface analytical tools in native-like bilayer environments. However, GPMVs do not easily rupture on conventional supports because of their high protein and cholesterol contents. Here, we demonstrate the possibility of using compression generated by the air-water interface to efficiently rupture GPMVs to form micron-sized supported membranes with native plasma membrane proteins. We demonstrated that not only lipid but also a native transmembrane protein in HeLa cells, Aquaporin 3 (AQP3), is mobile in the supported membrane platform. This convenient method for generating micron-sized supported membrane patches with mobile native transmembrane proteins could not only facilitate the study of membrane proteins by surface analytical tools, but could also enable us to use native membrane proteins for bio-sensing applications.
机译:能够直接获得具有天然膜蛋白的微米级细胞泡,巨大的质膜囊泡(GPMV),并将其沉积在平面支撑物上以形成支撑的质膜,这可以通过天然的各种表面分析工具研究膜蛋白类双层环境。但是,GPMV由于蛋白质和胆固醇含量高,因此在常规载体上不易破裂。在这里,我们证明了使用由空气-水界面产生的压缩来有效破裂GPMV形成具有天然质膜蛋白的微米级支持膜的可能性。我们证明,不仅脂质,而且HeLa细胞Aquaporin 3(AQP3)中的天然跨膜蛋白在支持的膜平台中均可移动。这种利用移动的天然跨膜蛋白生成微米级支持膜斑片的便捷方法,不仅可以促进通过表面分析工具对膜蛋白的研究,而且还使我们能够将天然膜蛋白用于生物传感应用。

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