Rondelli V, Fragneto G, Motta S, Del Favero E, Brocca P, Sonnino S, Cantù L
Dept. of Chemistry, University of Milan, Segrate, Italy.
Biochim Biophys Acta. 2012 Nov;1818(11):2860-7. doi: 10.1016/j.bbamem.2012.07.010. Epub 2012 Jul 22.
Neutron reflectivity has been applied to investigate different mixed asymmetric lipid systems, in the form of single "supported+floating" bilayers, made of phospholipids, cholesterol and GM1 ganglioside (Neu5Acα2-3(Galβ1-3GalNAcβ1-4)Galβ1-4Glcβ1Cer)) in bio-similar mole ratios. Bilayer preparation was carried out layer-by-layer with the Langmuir-Blodgett Langmuir-Schaefer techniques, allowing for compositional asymmetry in the system buildup. It is the first time that such a complex model membrane system is reported. Two important conclusions are drawn. First, it is experimentally shown that the presence of GM1 enforces an asymmetry in cholesterol distribution, opposite to what happens for a GM1-free membrane that, submitted to a similar procedure, results in a full symmetrization of cholesterol distribution. We underline that natural cholesterol has been used. Second, and most interesting, our results suggest that a preferential asymmetric distribution of GM1 and cholesterol is attained in a model membrane with biomimetic composition, revealing that a true coupling between the two molecular species occurs.
中子反射率已被用于研究不同的混合不对称脂质体系,其形式为单个“支撑+漂浮”双层膜,由磷脂、胆固醇和GM1神经节苷脂(Neu5Acα2-3(Galβ1-3GalNAcβ1-4)Galβ1-4Glcβ1Cer)以生物相似的摩尔比组成。双层膜的制备采用朗缪尔-布洛杰特(Langmuir-Blodgett)和朗缪尔-谢弗(Langmuir-Schaefer)技术逐层进行,从而在体系构建过程中实现组成的不对称性。这是首次报道这样一种复杂的模型膜系统。得出了两个重要结论。首先,实验表明,GM1的存在会导致胆固醇分布的不对称性,这与不含GM1的膜情况相反,后者在经过类似处理后,胆固醇分布会完全对称化。我们强调使用的是天然胆固醇。其次,也是最有趣的是,我们的结果表明,在具有仿生组成的模型膜中,GM1和胆固醇实现了优先的不对称分布,这表明这两种分子物种之间存在真正的耦合。