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. 2014 Nov;144(5):441–455. doi: 10.1085/jgp.201411184

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© 2014 Chowdhury et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

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Figure 4. — Interaction energies evaluated using GIA differ from those evaluated using FMC. (A–E) GIA was used to measure the interaction energies between A391-E395 (AE; A), A391-T469 (AT; B), A391-V476 (AV; C), E395-V476 (EV; D), and T469-V476 (TV; E). In each case the normalized Q-V curves of the single and double mutants were measured, from which the V_M was extracted and used to calculate the free energy of perturbation. The thermodynamic cycle for each pair is shown in the inset, in which each box corresponds to the WT or single or double mutants, colored as noted in the legends for each panel. (F) ΔΔG_GIA for each pair was calculated using the Q-V curves and compared with ΔΔG_FMC calculated using the Boltzmann fit parameters. For the ET pair, the horizontal green bar depicts ΔΔG_FMC evaluated under our experimental conditions. Error bars represent SEM.