Institute of Materials Research and Engineering, A∗STAR, Singapore, Singapore.
Biointerphases. 2011 Dec;6(4):153-7. doi: 10.1116/1.3644384.
Polymersomes are stable self-assembled architectures which mimic cell membranes. For characterization, membrane proteins can be incorporated into such bio-mimetic membranes by reconstitution methods, leading to so-called proteopolymersomes. In this work, we demonstrate the direct incorporation of a membrane protein into polymersome membranes by a cell-free expression system. Firstly, we demonstrate pore formation in the preformed polymersome membrane using α-hemolysin. Secondly, we use claudin-2, a protein involved in cell-cell interactions, to demonstrate the in vitro expression of a membrane protein into these polymersomes. Surface plasmon resonance (Biacore) binding studies with the claudin-2 proteopolymersomes and claudin-2 specific antibodies are performed to show the presence of the in vitro expressed protein in polymersome membranes.
聚合物囊泡是一种稳定的自组装结构，模拟细胞膜。为了进行表征，可以通过重建方法将膜蛋白掺入这种仿生膜中，从而得到所谓的蛋白聚合物囊泡。在这项工作中，我们通过无细胞表达系统证明了膜蛋白可以直接掺入聚合物囊泡膜中。首先，我们使用α-溶血素证明了预先形成的聚合物囊泡膜中的孔形成。其次，我们使用 Claudin-2，一种参与细胞间相互作用的蛋白质，证明了膜蛋白在体外表达到这些聚合物囊泡中。通过表面等离子体共振（Biacore）结合研究 Claudin-2 蛋白聚合物囊泡和 Claudin-2 特异性抗体，证明了体外表达的蛋白存在于聚合物囊泡膜中。
