Sensitive and High Resolution Localization and Tracking of Membrane Proteins in Live Cells with BRET

Tien Hung Lan, Qiuju Liu, Chunman Li, Guangyu Wu, Nevin A. Lambert

Research output: Contribution to journalArticle

44 Scopus citations

Abstract

Peripheral and integral membrane proteins can be located in several different subcellular compartments, and it is often necessary to determine the location of such proteins or to track their movement in living cells. Image-based colocalization of labeled membrane proteins and compartment markers is frequently used for this purpose, but this method is limited in terms of throughput and resolution. Here we show that bioluminescence resonance energy transfer (BRET) between membrane proteins of interest and compartment-targeted BRET partners can report subcellular location and movement of membrane proteins in live cells. The sensitivity of the method is sufficient to localize a few hundred protein copies per cell. The spatial resolution can be sufficient to determine membrane topology, and the temporal resolution is sufficient to track changes that occur in less than 1second. BRET requires little user intervention, and is thus amenable to large-scale experimental designs with standard instruments.

Original languageEnglish (US)
Pages (from-to)1450-1456
Number of pages7
JournalTraffic
Volume13
Issue number11
DOIs
StatePublished - Nov 2012

Keywords

  • Adrenergic
  • Bioluminescence
  • Colocalization
  • GTPase
  • Topology
  • Trafficking

ASJC Scopus subject areas

  • Structural Biology
  • Biochemistry
  • Molecular Biology
  • Genetics
  • Cell Biology

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