Department of Chemistry and Biochemistry and BIO5 Institute, University of Arizona, Tucson, Arizona 85721, USA.
J Am Chem Soc. 2010 May 26;132(20):7086-93. doi: 10.1021/ja100245d.
Suspended planar lipid membranes (or black lipid membranes (BLMs)) are widely used for studying reconstituted ion channels, although they lack the chemical and mechanical stability needed for incorporation into high-throughput biosensors and biochips. Lipid polymerization enhances BLM stability but is incompatible with ion channel function when membrane fluidity is required. Here, we demonstrate the preparation of a highly stable BLM that retains significant fluidity by using a mixture of polymerizable and nonpolymerizable phospholipids. Alamethicin, a voltage-gated peptide channel for which membrane fluidity is required for activity, was reconstituted into mixed BLMs prepared using bis-dienoyl phosphatidylcholine (bis-DenPC) and diphytanoyl phosphatidylcholine (DPhPC). Polymerization yielded BLMs that retain the fluidity required for alamethicin activity yet are stable for several days as compared to a few hours prior to polymerization. Thus, these polymerized, binary composition BLMs feature both fluidity and long-term stability.
悬浮平面脂质膜(或黑质脂质膜(BLM))广泛用于研究重组离子通道,尽管它们缺乏化学和机械稳定性,无法整合到高通量生物传感器和生物芯片中。脂质聚合增强了 BLM 的稳定性,但当需要膜流动性时,与离子通道功能不兼容。在这里,我们展示了一种高度稳定的 BLM 的制备方法,该 BLM 使用可聚合和不可聚合的磷脂混合物保持显著的流动性。alamethicin 是一种电压门控肽通道,其活性需要膜流动性,它被重新组装到使用双二烯酰基磷脂酰胆碱(bis-DenPC)和二植烷酰基磷脂酰胆碱(DPhPC)制备的混合 BLM 中。聚合生成的 BLM 保留了 alamethicin 活性所需的流动性,但与聚合前相比,稳定性可维持数天而非数小时。因此,这些聚合的二元组成 BLM 具有流动性和长期稳定性。