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合成并原位插入具有特异性荧光标记的膜蛋白到细胞大小的脂质体中。

Synthesis and in situ insertion of a site-specific fluorescently labeled membrane protein into cell-sized liposomes.

机构信息

Department of Bioscience and Biotechnology, Okayama University, Okayama 700-8530, Japan.

出版信息

Anal Biochem. 2011 Nov 1;418(1):97-101. doi: 10.1016/j.ab.2011.06.026. Epub 2011 Jun 26.

Abstract

The integral membrane protein bacteriorhodopsin, containing a fluorescent amino acid at a specific position, was synthesized in the presence of hydrated lipid films using an in vitro translation system expanded with a four-base codon/anticodon pair. Cell-sized liposomes with the labeled protein inserted into the liposome membranes were generated after the translation reaction. This study also demonstrated that this labeling method could be used to analyze the dynamic properties of membrane proteins in situ by fluorescence correlation spectroscopy.

摘要

含有特定荧光氨基酸的整合膜蛋白细菌视紫红质,在含有水合脂质膜的条件下,使用体外翻译系统,通过扩展的四碱基密码子/反密码子对进行合成。翻译反应后,生成了带有标记蛋白插入到脂质体膜中的细胞大小的脂质体。本研究还表明,这种标记方法可用于通过荧光相关光谱法原位分析膜蛋白的动态特性。

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