Measurement of molecular interactions in living cells by fluorescence resonance energy transfer between variants of the green fluorescent protein.

Journal Article (Journal Article)

Many signal transduction pathways operate through oligomerization of proteins into multi-subunit complexes. Although biochemical assays can identify potential protein-protein interactions, studying these interactions in living cells is more challenging. Fluorescence resonance energy transfer (FRET) has been used as a "spectroscopic ruler" to measure molecular proximity, but these methods have been limited by the need for chemical labeling of target proteins or labeled antibodies. We present methods for examining interactions between target proteins molecularly fused to cyan and yellow variants of the green fluorescent protein (GFP) by FRET in living cells. Flow cytometric and microscope-based methods are described that have been applied to a variety of interacting proteins.

Full Text

Duke Authors

Cited Authors

  • Siegel, RM; Chan, FK; Zacharias, DA; Swofford, R; Holmes, KL; Tsien, RY; Lenardo, MJ

Published Date

  • June 27, 2000

Published In

Volume / Issue

  • 2000 / 38

Start / End Page

  • pl1 -

PubMed ID

  • 11752595

Electronic International Standard Serial Number (EISSN)

  • 1525-8882

Digital Object Identifier (DOI)

  • 10.1126/stke.2000.38.pl1

Language

  • eng

Conference Location

  • United States