Deckmann M, Haimovitz R, Shinitzky M
Biochim Biophys Acta. 1985 Dec 5;821(2):334-40. doi: 10.1016/0005-2736(85)90103-8.
The overt effect of pressure on biological membranes is mediated predominantly through lipid condensation and disintegration of cytoskeletal polymers. These may lead to selective shedding of integral proteins, which could then be isolated by conventional means. In this study we have used the well characterised human erythrocyte membrane in order to establish the technical requirements for future use of pressure, as an alternative to detergents, in isolation of membrane proteins. Pressure of varying magnitude (300-1640 bar) and duration (5-60 min) was applied on human erythrocyte ghost membranes in suspension at different temperatures (4, 24 and 37 degrees C) and in the presence of various solutes. After ultracentrifugation protein and lipids remaining in the supernatant were quantified and analysed. It is indicated that selective integral membrane proteins can be shed off under defined conditions and presumably remain in solution by the support of strongly associated phospholipids and specific solutes. On the basis of our findings a series of technical recommendations for the isolation of specific membrane proteins is outlined.
压力对生物膜的明显影响主要通过脂质凝聚和细胞骨架聚合物的解体来介导。这些可能导致整合蛋白的选择性脱落，然后可以通过常规方法将其分离出来。在本研究中，我们使用了特性明确的人红细胞膜，以便确定未来使用压力（作为洗涤剂的替代品）分离膜蛋白的技术要求。在不同温度（4、24和37摄氏度）下，在各种溶质存在的情况下，对悬浮的人红细胞血影膜施加不同大小（300 - 1640巴）和持续时间（5 - 60分钟）的压力。超速离心后，对上清液中剩余的蛋白质和脂质进行定量和分析。结果表明，在特定条件下，选择性整合膜蛋白可以脱落，并且可能在强结合磷脂和特定溶质的支持下保留在溶液中。基于我们的研究结果，概述了一系列用于分离特定膜蛋白的技术建议。
