Three-chromophore FRET microscopy to analyze multiprotein interactions in living cells.

Emilia Galperin, Vladislav Verkhusha, Alexander Sorkin

Research output: Contribution to journalArticle

150 Citations (Scopus)

Abstract

Nearly every major process in a cell is carried out by assemblies of multiple dynamically interacting protein molecules. To study multi-protein interactions within such molecular machineries, we have developed a fluorescence microscopy method called three-chromophore fluorescence resonance energy transfer (3-FRET). This method allows analysis of three mutually dependent energy transfer processes between the fluorescent labels, such as cyan, yellow and monomeric red fluorescent proteins. Here, we describe both theoretical and experimental approaches that discriminate the parallel versus the sequential energy transfer processes in the 3-FRET system. These approaches were established in vitro and in cultured mammalian cells, using chimeric proteins consisting of two or three fluorescent proteins linked together. The 3-FRET microscopy was further applied to the analysis of three-protein interactions in the constitutive and activation-dependent complexes in single endosomal compartments. These data highlight the potential of 3-FRET microscopy in studies of spatial and temporal regulation of signaling processes in living cells.

Original languageEnglish (US)
Pages (from-to)209-217
Number of pages9
JournalCurrent Heart Failure Reports
Volume1
Issue number3
StatePublished - Dec 2004
Externally publishedYes

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Microscopy
Energy Transfer
Proteins
Fluorescence Resonance Energy Transfer
Fluorescence Microscopy
Cultured Cells

ASJC Scopus subject areas

  • Biotechnology
  • Molecular Biology
  • Cell Biology

Cite this

Three-chromophore FRET microscopy to analyze multiprotein interactions in living cells. / Galperin, Emilia; Verkhusha, Vladislav; Sorkin, Alexander.

In: Current Heart Failure Reports, Vol. 1, No. 3, 12.2004, p. 209-217.

Research output: Contribution to journalArticle

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