Direct preparation of giant proteo-liposomes by in vitro membrane protein synthesis
Direct preparation of giant proteo-liposomes by in vitro membrane protein synthesis
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DOI：
10.1016/j.jbiotec.2007.08.023
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发表时间：
2008-01-20
影响因子：
4.1
通讯作者：
Akiyoshi, Kazunari
中科院分区：
文献类型：
--
作者：
Nomura, Shin-ichiro M.;Kondoh, Satoshi;Akiyoshi, Kazunari
我们研究了在无细胞（体外）蛋白质合成中膜蛋白直接构成巨大脂质体。巨脂质体存在于小麦胚芽无细胞蛋白翻译系统的翻译反应混合物中。合成脱辅基细胞色素B（5）（M）及其融合蛋白，并直接定位于脂质体中。翻译反应后，通过简化的不连续密度梯度离心分离蛋白脂质体。以同样的方法合成了脱辅基细胞色素b5结合的二氢叶酸还原酶（DHFR），并将其直接展示在脂质体表面。B5作为“疏水标签”用于募集到脂质体表面。(c)2007 Elsevier B.V.保留所有权利。
We investigated the direct constitution of membrane proteins into giant liposomes in cell-free (in vitro) protein synthesis. Giant liposomes were present in a translation reaction cocktail of a wheat germ cell-free protein translation system. Apo cytochrome b(5) (M) and its fusion proteins were synthesized and directly localized in the liposomes. After the translation reaction, the proteo-liposomes were isolated by simplified discontinuous density-gradient centrifugation. Apo cytochrome b5 conjugated dihydrofolate reductase (DHFR) was synthesized in the same procedure and the protein was directly displayed on the liposome surface. b5 acts as a "hydrophobic tag" for recruitment to the liposome surface. (c) 2007 Elsevier B.V. All rights reserved.