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

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acids / chemistry
  • Bacteriorhodopsins / chemistry*
  • Bacteriorhodopsins / genetics
  • Codon
  • Fluorescent Dyes / chemistry
  • Liposomes / chemistry*
  • Microscopy, Confocal
  • Mutagenesis, Insertional
  • RNA, Messenger / genetics
  • RNA, Transfer / genetics
  • Spectrometry, Fluorescence / methods*

Substances

  • Amino Acids
  • Codon
  • Fluorescent Dyes
  • Liposomes
  • RNA, Messenger
  • Bacteriorhodopsins
  • RNA, Transfer