Division of Biophysics, Department of Biology, University of Osnabrück, 49069 Osnabrück, Germany.
J Am Chem Soc. 2013 Jan 30;135(4):1189-92. doi: 10.1021/ja310186g. Epub 2013 Jan 16.
We have established an approach for the spatial control of lipid phase separation in tethered polymer-supported membranes (PSMs), which were obtained by vesicle fusion on a poly(ethylene glycol) polymer brush functionalized with fatty acid moieties. Phase separation of ternary lipid mixtures (1,2-dioleoyl-sn-glycero-3-phosphocholine/sphingomyelin/cholesterol) into liquid-disordered (l(d)) and liquid-ordered (l(o)) phases within both leaflets was obtained with palmitic acid as the anchoring group. In contrast, tethering of the PSM with oleic acid interfered with the phase separation in the surface-proximal leaflet. We exploited this feature for the assembly of l(o) domains within PSMs into defined structures by binary micropatterning of palmitic and oleic acid into complementary areas. Ternary lipid mixtures spontaneously separated into l(o) and l(d) phases controlled by the geometry of the underlying tethers. Transmembrane proteins reconstituted in these phase-separated PSMs strictly partitioned into the l(d) phase. Hence, the l(o) phase could be used for confining transmembrane proteins into microscopic and submicroscopic domains.
我们已经建立了一种方法来控制连接聚合物支撑膜(PSM)中的脂相分离,这些膜是通过在脂肪酸部分官能化的聚乙二醇聚合物刷上的囊泡融合获得的。通过将三种脂质混合物(1,2-二油酰基-sn-甘油-3-磷酸胆碱/鞘磷脂/胆固醇)相分离成双层中的无序相(l(d))和有序相(l(o)),其中棕榈酸作为锚定基团。相比之下,PSM 与油酸的连接干扰了表面近侧单层中的相分离。我们利用这一特性,通过将棕榈酸和油酸二元微图案化成互补区域,将 PSM 内的 l(o)域组装成特定结构。三元脂质混合物自发地分离成 l(o)和 l(d)相,由底层连接物的几何形状控制。再嵌入这些相分离 PSM 中的跨膜蛋白严格分配到 l(d)相中。因此,l(o)相可用于将跨膜蛋白限制在微观和亚微观域内。