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Synthesis and in situ insertion of a site-specific fluorescently labeled membrane protein into cell-sized liposomes |
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| Authors: | Takuma Ohtsuka Satoshi Neki Tamotsu Kanai Kazunari Akiyoshi Shin-ichiro M. Nomura Takashi Ohtsuki |
| Affiliation: | aDepartment of Bioscience and Biotechnology, Okayama University, Okayama 700-8530, Japan;bDepartment of Synthetic Chemistry and Biological Chemistry, Kyoto University, Kyoto 606-8501, Japan;cDepartment of Polymer Chemistry, Kyoto University, Kyoto 606-8501, Japan;dInstitute for Integrated Cell Material Sciences (iCeMS), Kyoto University, Kyoto 606-8501, Japan;ePrecursory Research for Embryonic Science and Technology (PRESTO), Japan Science and Technology Agency (JST), Saitama 332-0012, Japan |
| 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. |
| Keywords: | Protein labeling Membrane protein Liposome Fluorescence correlation spectroscopy Four-base codon Translation |
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