title: Synthesis and in situ insertion of a site-specific fluorescently labeled membrane protein into cell-sized liposomes.
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.
authors.affiliation: Department of Bioscience and Biotechnology, Okayama University, Okayama 700-8530, Japan.
authors.forename: Takuma
authors.initials: T
authors.lastname: Ohtsuka
authors.forename: Satoshi
authors.initials: S
authors.lastname: Neki
authors.forename: Tamotsu
authors.initials: T
authors.lastname: Kanai
authors.forename: Kazunari
authors.initials: K
authors.lastname: Akiyoshi
authors.forename: Shin-ichiro M
authors.initials: SM
authors.lastname: Nomura
authors.forename: Takashi
authors.initials: T
authors.lastname: Ohtsuki
mesh_terms: D000596:Amino Acids / Q000737:chemistry; D001436:Bacteriorhodopsins / Q000737:chemistry* / Q000235:genetics; D003062:Codon; D005456:Fluorescent Dyes / Q000737:chemistry; D008081:Liposomes / Q000737:chemistry*; D018613:Microscopy, Confocal; D016254:Mutagenesis, Insertional; D012333:RNA, Messenger / Q000235:genetics; D012343:RNA, Transfer / Q000235:genetics; D013050:Spectrometry, Fluorescence / Q000379:methods*
chemical_list: D000596:Amino Acids; D003062:Codon; D005456:Fluorescent Dyes; D008081:Liposomes; D012333:RNA, Messenger; D001436:Bacteriorhodopsins; D012343:RNA, Transfer