Formation of Giant Protein Vesicles by a Lipid Cosolvent Method
Formation of Giant Protein Vesicles by a Lipid Cosolvent Method
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DOI：
10.1002/cbic.201100537
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发表时间：
2011-12-16
期刊：
影响因子：
3.2
通讯作者：
Helix-Nielsen, Claus
中科院分区：
文献类型：
--
作者：
Hansen, Jesper S.;Vararattanavech, Ardcharaporn;Helix-Nielsen, Claus
本文介绍了一种利用大囊泡（0.10.2 μ m）与重组膜蛋白进行溶剂驱动融合制备>= 10 μ m巨蛋白囊泡（GPVs）的方法。我们发现GPVs的形成是由蛋白质重组的大单层囊泡（LUVs）与含脂溶剂相的旋转混合进行的。我们以正癸烷和角鲨烯为溶剂制备了GPVs，并用极性敏感探针“巴丹”标记水通道蛋白跨膜区周围的极性进行了广义极化成像（GP）监测。具体来说，我们创建了菠菜SoPIP2的gpv；1和大肠杆菌AqpZ水通道蛋白。我们的研究结果表明，形成的gpv双分子层内的疏水相互作用不仅受溶剂分配倾向的影响，还受脂质组成和膜蛋白异构体的影响。
This paper describes a method to create giant protein vesicles (GPVs) of >= 10 mu m by solvent-driven fusion of large vesicles (0.10.2 mu m) with reconstituted membrane proteins. We found that formation of GPVs proceeded from rotational mixing of protein-reconstituted large unilamellar vesicles (LUVs) with a lipid-containing solvent phase. We made GPVs by using n-decane and squalene as solvents, and applied generalized polarization (GP) imaging to monitor the polarity around the protein transmembrane region of aquaporins labeled with the polarity-sensitive probe Badan. Specifically, we created GPVs of spinach SoPIP2;1 and E. coli AqpZ aquaporins. Our findings show that hydrophobic interactions within the bilayer of formed GPVs are influenced not only by the solvent partitioning propensity, but also by lipid composition and membrane protein isoform.