Epidermal growth factor receptor dimerization monitored in live cells.

B T Blakely; F M Rossi; B Tillotson; M Palmer; A Estelles; H M Blau

doi:10.1038/72686

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Epidermal growth factor receptor dimerization monitored in live cells.

B T Blakely·F M Rossi·B Tillotson·M Palmer·A Estelles·H M Blau

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Abstract

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.

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Animals Cell Membrane Dimerization Epidermal Growth Factor ErbB Receptors Genetic Complementation Test Humans Mice Protein Binding Recombinant Fusion Proteins beta-Galactosidase Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

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B T Blakely, F M Rossi, B Tillotson, M Palmer, A Estelles, H M Blau. 2000. Epidermal growth factor receptor dimerization monitored in live cells.. https://doi.org/10.1038/72686

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