Department of Chemical and Biological Engineering, University of Wisconsin-Madison, 1415 Engineering Drive, Madison, Wisconsin 53706, USA.
Langmuir. 2012 Apr 17;28(15):6364-76. doi: 10.1021/la300108f. Epub 2012 Apr 3.
We report that specific binding of ligand-functionalized (biotinylated) phospholipid vesicles (diameter = 120 ± 19 nm) to a monolayer of proteins (streptavidin or anti-biotin antibody) adsorbed at an interface between an aqueous phase and an immiscible film of a thermotropic liquid crystal (LC) [nematic 4'-pentyl-4-cyanobiphenyl (5CB)] triggers a continuous orientational ordering transition (continuous change in the tilt) in the LC. Results presented in this paper indicate that, following the capture of the vesicles at the LC interface via the specific binding interaction, phospholipids are transferred from the vesicles onto the LC interface to form a monolayer, reorganizing and partially displacing proteins from the LC interface. The dynamics of this process are accelerated substantially by the specific binding event relative to a protein-decorated interface of a LC that does not bind the ligands presented by the vesicles. The observation of the continuous change in the ordering of the LC, when combined with other results presented in this paper, is significant, as it is consistent with the presence of suboptical domains of proteins and phospholipids on the LC interface. An additional significant hypothesis that emerges from the work reported in this paper is that the ordering transition of the LC is strongly influenced by the bound state of the protein adsorbed on the LC interface, as evidenced by the influence on the LC of (i) "crowding" of the protein within a monolayer formed at the LC interface and (ii) aging of the proteins on the LC interface. Overall, these results demonstrate that ordering transitions in LCs can be used to provide fundamental insights into the competitive adsorption of proteins and lipids at oil-water interfaces and that LC ordering transitions have the potential to be useful for reporting specific binding events involving vesicles and proteins.
我们报告说,配体功能化(生物素化)的磷脂囊泡(直径= 120 ± 19nm)与吸附在水相和热致液晶(向列相 4'-戊基-4-氰基联苯(5CB))不混相膜之间界面上的蛋白质(链霉亲和素或抗生物素抗体)单层的特异性结合触发了液晶的连续取向有序转变(倾斜连续变化)。本文介绍的结果表明,在通过特异性结合相互作用将囊泡捕获到 LC 界面之后,磷脂从囊泡转移到 LC 界面以形成单层,重组并部分从 LC 界面置换蛋白质。与不结合囊泡提供的配体的 LC 上的蛋白质修饰界面相比,该过程的动力学通过特异性结合事件大大加速。观察到 LC 的有序性的连续变化,与本文中提出的其他结果相结合,是非常重要的,因为它与 LC 界面上的蛋白质和磷脂的亚光域的存在一致。从本文报道的工作中得出的另一个重要假设是,吸附在 LC 界面上的蛋白质的结合状态强烈影响 LC 的有序转变,这可以通过 LC 上的以下两种情况来证明:(i)在 LC 界面上形成的单层中蛋白质的“拥挤”,以及(ii)LC 界面上蛋白质的老化。总体而言,这些结果表明,LC 的有序转变可用于深入了解油-水界面上蛋白质和脂质的竞争吸附,并且 LC 有序转变有可能用于报告涉及囊泡和蛋白质的特异性结合事件。