Department of Biology, University of Pennsylvania, Philadelphia, PA 19104-6313.
Department of Genetics, Rutgers University, Piscataway, NJ 08854.
Proc Natl Acad Sci U S A. 2019 Jan 15;116(3):744-752. doi: 10.1073/pnas.1811307116. Epub 2018 Dec 27.
Cell-like hybrids from natural and synthetic amphiphiles provide a platform to engineer functions of synthetic cells and protocells. Cell membranes and vesicles prepared from human cell membranes are relatively unstable in vitro and therefore are difficult to study. The thicknesses of biological membranes and vesicles self-assembled from amphiphilic Janus dendrimers, known as dendrimersomes, are comparable. This feature facilitated the coassembly of functional cell-like hybrid vesicles from giant dendrimersomes and bacterial membrane vesicles generated from the very stable bacterial cell after enzymatic degradation of its outer membrane. Human cells are fragile and require only mild centrifugation to be dismantled and subsequently reconstituted into vesicles. Here we report the coassembly of human membrane vesicles with dendrimersomes. The resulting giant hybrid vesicles containing human cell membranes, their components, and Janus dendrimers are stable for at least 1 y. To demonstrate the utility of cell-like hybrid vesicles, hybrids from dendrimersomes and bacterial membrane vesicles containing YadA, a bacterial adhesin protein, were prepared. The latter cell-like hybrids were recognized by human cells, allowing for adhesion and entry of the hybrid bacterial vesicles into human cells in vitro.
天然和合成两亲物形成的类细胞杂种为工程化合成细胞和原细胞的功能提供了一个平台。用人细胞膜制备的细胞膜和囊泡在体外相对不稳定,因此难以研究。由两亲性 Janus 树状大分子自组装形成的生物膜和囊泡的厚度与细胞膜相当。这一特性促进了功能型类细胞杂种囊泡的共组装,这些囊泡由巨大的树状大分子囊泡和细菌膜囊泡组成,细菌膜囊泡是通过酶降解其外膜从非常稳定的细菌细胞中产生的。人细胞很脆弱,只需轻微离心即可被拆开,然后重新组装成囊泡。在这里,我们报告了用人细胞膜囊泡与树状大分子囊泡的共组装。所得的含有人类细胞膜、其成分和 Janus 树状大分子的巨大杂交囊泡至少稳定 1 年。为了证明类细胞杂种囊泡的实用性,制备了含有细菌黏附蛋白 YadA 的树状大分子囊泡和细菌膜囊泡的杂种。后者的类细胞杂种被人细胞识别,允许杂交细菌囊泡进入人细胞的黏附和进入。