Figure 5.

Histograms of ΔΔG_bind for single and double BPTI mutants. Single BPTI mutants interacting with (A) BT; (B) ChT; and (C) MT. Double BPTI mutants interacting with (D) BT; (E) ChT; and (F) MT. Dark red represents the actual measured values, while light red represents the values that would result from all single mutations being additive. Mean value for ΔΔG_bind for each histogram is displayed on top of each graph; in parentheses is the value that would result from all double mutations being additive. While all 228 single mutants are incorporated into the single mutant histograms for all proteases, only ∼50% of double mutants are summarized. The data are available in the Source Data file in the Supporting Information.