Development of cell-free synthesis and imaging of membrane proteins in giant vesicles

巨囊泡膜蛋白无细胞合成和成像的进展

• 批准号: 21750073
• 负责人: TOYOTA Taro
• 金额: $ 3万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Young Scientists (B)
• 财政年份: 2009
• 资助国家: 日本
• 起止时间: 2009 至 2010
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-21750073/
• 关键词: 生体分析; 膜タンパク質; ベシクル; 反射干渉顕微鏡; 干渉顕微鏡

The aim of this study is the development of an analytical method to observe the non-equilibrium dynamics of membrane proteins incorporated into model cell membrane. Giant vesicle (GV), which is closed bilayer lipid membrane with cellular size, was used as a reactor of model cell membrane for the cell-free synthesis of a fluorescent membrane protein, eGFP-BmOR1, and the spontaneous membrane insertion of eGFP-BmOR1 synthesized inside of GV was successfully observed. Upon the construction of a reflective interference contrast microscope, the non-equilibrium dynamics of GV settling on a glass substrate was revealed in a nanometer thickness resolution. The combination of these two technologies is expected to promote further progress in the analysis of the non-equilibrium dynamics of membrane proteins and lipid membrane.

本研究的目的是开发一种分析方法来观察模型细胞膜中膜蛋白的非平衡动力学。巨型囊泡（GV）是一种具有细胞大小的封闭双层脂质膜，被用作模型细胞膜反应器，用于荧光膜蛋白 eGFP-BmOR1 的无细胞合成，以及 eGFP-BmOR1 的自发膜插入成功观察到GV内部合成。在构建反射干涉对比显微镜后，以纳米厚度分辨率揭示了玻璃基板上 GV 沉降的非平衡动力学。这两项技术的结合有望促进膜蛋白和脂质膜非平衡动力学分析的进一步进展。

项目成果

Population Analysis of Structural Properties of Giant Liposomes by Flow Cytometry

• DOI: 10.1021/la902237y
• 发表时间: 2009-09-15
• 期刊: LANGMUIR
• 影响因子: 3.9
• 作者: Nishimura, Kazuya;Hosoi, Tomohiro;Yomo, Tetsuya
• 通讯作者: Yomo, Tetsuya