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

Takuma Ohtsuka, Satoshi Neki, Tamotsu Kanai, Kazunari Akiyoshi, Shin Ichiro M Nomura, Takashi Ohtsuki

Research output: Contribution to journalArticle

9 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)97-101
Number of pages5
JournalAnalytical Biochemistry
Volume418
Issue number1
DOIs
Publication statusPublished - Nov 1 2011

Fingerprint

Liposomes
Membrane Proteins
Bacteriorhodopsins
Anticodon
Fluorescence Spectrometry
Codon
Labeling
Fluorescence
Spectroscopy
Membranes
Lipids
Amino Acids
Proteins
In Vitro Techniques

Keywords

  • Fluorescence correlation spectroscopy
  • Four-base codon
  • Liposome
  • Membrane protein
  • Protein labeling
  • Translation

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology
  • Cell Biology

Cite this

Synthesis and in situ insertion of a site-specific fluorescently labeled membrane protein into cell-sized liposomes. / Ohtsuka, Takuma; Neki, Satoshi; Kanai, Tamotsu; Akiyoshi, Kazunari; Nomura, Shin Ichiro M; Ohtsuki, Takashi.

In: Analytical Biochemistry, Vol. 418, No. 1, 01.11.2011, p. 97-101.

Research output: Contribution to journalArticle

Ohtsuka, Takuma ; Neki, Satoshi ; Kanai, Tamotsu ; Akiyoshi, Kazunari ; Nomura, Shin Ichiro M ; Ohtsuki, Takashi. / Synthesis and in situ insertion of a site-specific fluorescently labeled membrane protein into cell-sized liposomes. In: Analytical Biochemistry. 2011 ; Vol. 418, No. 1. pp. 97-101.
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