Lujan Neutron Scattering Center, Los Alamos National Laboratory, Los Alamos, New Mexico, USA.
Biophys J. 2010 Sep 8;99(5):1475-81. doi: 10.1016/j.bpj.2010.06.028.
The interaction of beta-cyclodextrin (beta-CD) with mixed bilayers composed of sphingomylein and cholesterol (Chol) above and below the accepted stable complexation ratio (67:33) was investigated. Membranes with the same (symmetric) and different (asymmetric) compositions in their inner and outer leaflets were deposited at surface pressures of 20, 30, and 40 mN/m at the solid-liquid interface. Using neutron reflectometry, membranes of various global molar ratios (defined as the sum of the molar ratios of the inner and outer leaflets), were characterized before and after beta-CD was added to the subphase. The structure of bilayers with global molar ratios at or above the stable complexation ratio was unchanged by beta-CD, indicating that beta-CD is unable to remove sphingomyelin or complexed Chol. However, beta-CD removed all uncomplexed Chol from bilayers composed of global molar ratios below the stable complexation ratio. The removal of Chol by beta-CD was independent of the initial structure of the membranes as deposited, suggesting that asymmetric membranes homogenize by the exchange of molecules between leaflets. The interaction of beta-CD with the aforementioned membranes was independent of the deposition surface pressure except for a symmetric 50:50 membrane deposited at 40 mN/m. The scattering from 50:50 bilayers with higher packing densities (deposited at 40 mN/m) was unaffected by beta-CD, suggesting that the removal of Chol can depend on both the composition and packing density of the membrane.
研究了β-环糊精(β-CD)与鞘磷脂和胆固醇(Chol)混合双层的相互作用,这些双层的组成比例高于且低于公认的稳定配合比(67:33)。在固-液界面上,将具有相同(对称)和不同(不对称)内层和外层组成的双层以 20、30 和 40 mN/m 的表面压力沉积。使用中子反射测量法,在亚相中添加β-CD 之前和之后,对各种总摩尔比(定义为内层和外层摩尔比之和)的膜进行了表征。总摩尔比等于或高于稳定配合比的双层结构不受β-CD 的影响,这表明β-CD 无法去除鞘磷脂或与之复合的 Chol。然而,β-CD 从总摩尔比低于稳定配合比的双层中去除了所有未复合的 Chol。β-CD 对 Chol 的去除与膜的初始结构无关,这表明不对称膜通过分子在双层之间的交换而均匀化。β-CD 与上述膜的相互作用除了在 40 mN/m 下沉积的对称 50:50 膜外,与沉积表面压力无关。具有更高堆积密度(在 40 mN/m 下沉积)的 50:50 双层的散射不受β-CD 的影响,这表明 Chol 的去除可能取决于膜的组成和堆积密度。