Epidermal growth factor receptor dimerization monitored in live cells

Nat Biotechnol. 2000 Feb;18(2):218-22. doi: 10.1038/72686.


We present a method for monitoring receptor dimerization at the membrane of live cells. Chimeric proteins containing the epidermal growth factor (EGF) receptor extracellular and transmembrane domains fused to weakly complementing beta-galactosidase (beta-gal) deletion mutants were expressed in cells in culture. Treatment of the cells with EGF-like compounds for as little as 15 s resulted in chimeric receptor dimerization detectable as beta-gal enzymatic activity. The dose response of chimeric receptors was ligand specific. beta-galactosidase complementation was reversible upon removal of ligand and could be reinduced. Antibodies that block ligand binding inhibited receptor dimerization and beta-gal complementation. These results demonstrate that beta-gal complementation provides a rapid, simple, and sensitive assay for protein interactions and for detecting and monitoring the kinetics of receptor dimerization.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Cell Membrane / metabolism
  • Dimerization
  • Epidermal Growth Factor / pharmacology*
  • ErbB Receptors / drug effects
  • ErbB Receptors / genetics
  • ErbB Receptors / metabolism*
  • Genetic Complementation Test
  • Humans
  • Mice
  • Protein Binding
  • Recombinant Fusion Proteins / metabolism
  • beta-Galactosidase / genetics
  • beta-Galactosidase / metabolism


  • Recombinant Fusion Proteins
  • Epidermal Growth Factor
  • ErbB Receptors
  • beta-Galactosidase