Department of Chemistry, Stanford University, Stanford, California 94305-5080, USA.
Langmuir. 2010 Jun 1;26(11):8666-72. doi: 10.1021/la904822f.
Specific membrane interactions such as lipid vesicle docking and fusion can be mediated by synthetic DNA-lipid conjugates as a model for the protein-driven processes that are ubiquitous in biological systems. Here we present a method of tethering vesicles to a supported lipid bilayer that allows the simultaneous deposition of cognate vesicle partners displaying complementary DNA, resulting in well-mixed populations of tethered vesicles that are laterally mobile. Vesicles are covalently attached to a supporting lipid bilayer using a DNA-templated click reaction; then DNA-mediated interactions between tethered vesicles are triggered by spiking the salt concentration. These interactions, such as docking and fusion, can then be observed for individual vesicles as they collide on the surface. The architecture of this new system also permits control over the number of lipid anchors that tether the vesicle to the supporting bilayer. The diffusion coefficient of tethered vesicles anchored by two lipids is approximately 1.6-fold slower than that of vesicles anchored only with a single lipid, consistent with a simple physical model.
特定的膜相互作用,如脂质囊泡的对接和融合,可以通过合成 DNA-脂质缀合物来介导,作为蛋白质驱动过程的模型,这些过程在生物系统中普遍存在。在这里,我们提出了一种将囊泡固定在支撑脂质双层上的方法,该方法允许同时沉积显示互补 DNA 的同源囊泡伴侣,从而形成混合良好的、侧向可移动的固定化囊泡群体。囊泡通过 DNA 模板点击反应共价连接到支撑脂质双层上;然后通过提高盐浓度来触发囊泡之间的 DNA 介导相互作用。这些相互作用,如对接和融合,可以在单个囊泡在表面上碰撞时进行观察。该新系统的结构还允许控制将囊泡固定到支撑双层的脂质锚的数量。用两个脂质锚定的固定化囊泡的扩散系数比只用一个脂质锚定的囊泡大约慢 1.6 倍,与简单的物理模型一致。