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. Author manuscript; available in PMC: 2013 Jan 14.
Published in final edited form as: J Biol Chem. 2012 Mar 22;287(21):17459–17470. doi: 10.1074/jbc.M111.331967

FIG. 1.

FIG. 1

The mRFP-IgEFc-eGFP biosensor reveals IgEFc to be bent in solution and to undergo conformational change on binding to sFcεRIα. A: Cartoons of the three fluorescent constructs. B: Steady-state fluorescence excitation scan (emission detected at 610nm) of either the FRET biosensor mRFP-IgEFc-eGFP (blue line), or control molecules mRFP-IgEFc alone (red dashed line), or with IgEFc-eGFP (cyan line). The increase in fluorescence shown here is indicative of energy transfer. C: Cartoon of the proposed conformational change model to be tested using the mRFP-IgEFc-eGFP biosensor. D: Steady-state fluorescence excitation scan (emission detected at 610nm) of either the FRET biosensor mRFP-IgEFc-eGFP alone (blue line) or saturated with sFcεRIα (green line), or control molecules mRFP-IgEFc and IgEFc-eGFP alone (cyan line) or with sFcεRIα (black dashed line). The fluorescence increases when sFcεRIα binds to the mRFP-IgEFc-GFP biosensor indicative of additional bending within IgEFc.