Rapid preparation of giant unilamellar vesicles
Rapid preparation of giant unilamellar vesicles
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DOI：
10.1073/pnas.93.21.11443
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发表时间：
1996-10-15
影响因子：
11.1
通讯作者：
Zare, RN
中科院分区：
文献类型：
--
作者：
Moscho, A;Orwar, O;Zare, RN
我们在这里报告了一种快速蒸发方法，在高产量的巨型单层囊泡直径达50 μ m。囊泡仅在2分钟后获得，并且可以由不同的磷脂制备，包括L-α-磷脂酰胆碱（卵磷脂）、二棕榈油酰L-α-磷脂酰胆碱和β-花生四烯酸酰γ-棕榈酰L-α-磷脂酰胆碱。囊泡可以在蒸馏水和Hepes，磷酸盐和berate缓冲液中产生，pH范围为7.0至11.5，离子强度高达50 mM。短的制备时间允许封装不稳定的分子靶点或具有高催化活性的酶。已经制备了细胞大小的蛋白脂质体，其中γ-谷氨酰转移酶（EC 2.3.2.2）功能性地并入膜壁中。
We report here a rapid evaporation method that produces in high yield giant unilamellar vesicles up to 50 mu m in diameter. The vesicles are obtained after only 2 min and can be prepared from different phospholipids, including L-alpha-phosphatidylcholine (lecithin), dipalmitoleoyl L-alpha-phosphatidylcholine, and beta-arachidonoyl gamma-palmitoyl L-alpha-phosphatidylcholine. Vesicles can be produced in distilled water and in Hepes, phosphate, and berate buffers in the pH range of 7.0 to 11.5 with ionic strengths up to 50 mM. The short preparation time allows encapsulation of labile molecular targets or enzymes with high catalytic activities. Cell-sized proteoliposomes have been prepared in which gamma-glutamyltransferase (EC 2.3.2.2) was functionally incorporated into the membrane wall.