Special Section on Visible Fluorescent Proteins

Dimerization between aequorea fluorescent proteins does not affect interaction between tagged estrogen receptors in living cells

[+] Author Affiliations
Eric M. Kofoed, Martin Guerbadot, Fred Schaufele

University of California, San Francisco, Diabetes Center and Department of Medicine, S-1230, 513 Parnassus, San Francisco, California 94143-0540

J. Biomed. Opt. 13(3), 031207 (June 23, 2008). doi:10.1117/1.2940366
History: Received August 06, 2007; Revised January 23, 2008; Accepted January 24, 2008; Published June 23, 2008
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Förster resonance energy transfer (FRET) detection of protein interaction in living cells is commonly measured following the expression of interacting proteins genetically fused to the cyan (CFP) and yellow (YFP) derivatives of the Aequorea victoria fluorescent protein (FP). These FPs can dimerize at mM concentrations, which may introduce artifacts into the measurement of interaction between proteins that are fused with the FPs. Here, FRET analysis of the interaction between estrogen receptors (alpha isoform, ERα) labeled with “wild-type” CFP and YFP is compared with that of ERα labeled with “monomeric” A206K mutants of CFP and YFP. The intracellular equilibrium dissociation constant for the hormone-induced ERα-ERα interaction is similar for ERα labeled with wild-type or monomeric FPs. However, the measurement of energy transfer measured for ERα-ERα interaction in each cell is less consistent with the monomeric FPs. Thus, dimerization of the FPs does not affect the kinetics of ERα-ERα interaction but, when brought close together via ERα-ERα interaction, FP dimerization modestly improves FRET measurement.

Figures in this Article
© 2008 Society of Photo-Optical Instrumentation Engineers

Citation

Eric M. Kofoed ; Martin Guerbadot and Fred Schaufele
"Dimerization between aequorea fluorescent proteins does not affect interaction between tagged estrogen receptors in living cells", J. Biomed. Opt. 13(3), 031207 (June 23, 2008). ; http://dx.doi.org/10.1117/1.2940366


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