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Asymmetric Structural Features in Single Supported Lipid Bilayers Containing Cholesterol and GM1 Resolved with Synchrotron X-Ray Reflectivity

机译：用同步辐射X射线反射率解析的含胆固醇和GM1的单支撑脂质双层的不对称结构特征

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The cell membrane comprises numerous protein and lipid molecules capable of asymmetric organization between leaflets and liquid-liquid phase separation. We use single supported lipid bilayers (SLBs) to model cell membranes, and study how cholesterol and asymmetrically oriented ganglioside receptor GM1 affect membrane structure using synchrotron x-ray reflectivity. Using mixtures of cholesterol, sphingomyelin, and 1,2-dioleoyl-sn-glycero-3-phosphocholine, we characterize the structure of liquid-ordered and liquid-disordered SLBs in terms of acyl-chain density, headgroup size, and leaflet thickness. SLBs modeling the liquid-ordered phase are 10 Å thicker and have a higher acyl-chain electron density (〈ρchain〉 = 0.33 e−/Å3) compared to SLBs modeling the liquid-disordered phase, or pure phosphatidylcholine SLBs (〈ρchain〉 = 0.28 e−/Å3). Incorporating GM1 into the distal bilayer leaflet results in membrane asymmetry and thickening of the leaflet of 4–9 Å. The structural effect of GM1 is more complex in SLBs of cholesterol/sphingomyelin/1,2-dioleoyl-sn-glycero-3-phosphocholine, where the distal chains show a high electron density (〈ρchain〉 = 0.33 e−/Å3) and the lipid diffusion constant is reduced by ∼50%, as measured by fluorescence microscopy. These results give quantitative information about the leaflet asymmetry and electron density changes induced by receptor molecules that penetrate a single lipid bilayer.
机译：细胞膜包含许多蛋白质和脂质分子，能够在小叶之间进行不对称组织和液-液相分离。我们使用单个支持的脂质双层（SLBs）来建模细胞膜，并使用同步加速器X射线反射率研究胆固醇和不对称取向的神经节苷脂受体GM1如何影响膜结构。使用胆固醇，鞘磷脂和1,2-二油酰基-sn-甘油-3-磷酸胆碱的混合物，我们根据酰基链密度，头基团大小和小叶厚度表征了液体有序和液体无序的SLB的结构。与模拟液相无序相的SLB或纯磷脂酰胆碱SLB相比，模拟液相有序相的SLB厚10Å，并且具有更高的酰基链电子密度（〈ρ链〉 = 0.33 e- /Å3）。 0.28 e- /Å3）。将GM1掺入远端双层小叶会导致膜不对称，并使小叶增厚4–9Å。 GM1的结构效应在胆固醇/鞘磷脂/ 1,2-油酰-sn-甘油-3-磷酸胆碱的SLB中更为复杂，其中远端链显示出高电子密度（〈ρ链〉 = 0.33 e- /Å3），并且通过荧光显微镜观察，脂质的扩散常数降低了约50％。这些结果提供了有关渗透单个脂质双层的受体分子诱导的小叶不对称性和电子密度变化的定量信息。

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