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FIGURE 4. Double mutant cycle analysis. A, glutamate-gated currents at oocytes expressing double mutant T93S/R95K or R76K/T93S receptors and mean (± S.E.) data. Robust activation by IVM indicates successful expression of R76K/T93S despite small responses to glutamate. WT and single mutant data are repeated from earlier figures for comparison. B, principles of double mutant cycle analysis and analysis of Arg76–Thr93 and Arg95–Thr93 coupling. For two residues “A” and “B,” EC50 values of WT (AB), single mutant A′B, single mutant AB′, and double mutant A′B′ are used to calculate the coupling coefficient, Ω, from which the coupling energy, ΔΔG, can be calculated (22). Our final values for Ω and ΔΔG are only estimates because the EC50 values of certain single and double mutants could not be calculated: here the EC50 values have been estimated from A. Image published in: Lynagh T et al. (2017) © 2017 by The American Society for Biochemistry and Molecular Biology, Inc. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View

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