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Fig. S1. Correlation between 109 measured 1DNH residual dipolar couplings of EW29Ch in the sugar-free state and those calculated from the crystal structure of lactose-bound EW29Ch.

Fig. S2. Plots of the 15N relaxation data [R1(15N), R2(15N) and 15N{1H}-NOE] for the backbone of EW29Ch in the sugar-free state and in the lactose-bound states at [EW29Ch]/[lactose] = 1/1, 1/8 and 1/80.

Fig. S3. Mapping of the transverse relaxation rate (R2 s−1) differences (ΔR2) between R2 in the sugar-free state (R2-free s−1) and those in the lactose-bound states (R2-bound s−1) at [EW29Ch]/[lactose] = 1/1, 1/8 or 1/80 (ΔR2 = R2-free − R2-bound) on a ribbon diagram of the sugar-binding sites of the lowest-energy NMR structure of sugar-free EW29Ch in subdomain α (left), subdomain β (centre) and subdomain γ (right).

Fig. S4. Mapping of the transverse relaxation rate (R2 s−1) differences (ΔR2) between R2 in the sugar-free states (R2-free s−1) and the lactose-bound state (R2-bound s−1) at [EW29Ch]/[lactose] = 1/1, 1/8 or 1/80 (ΔR= R2-free − R2-bound) on a ribbon diagram of the sugar-binding sites of the crystal structure of lactose-bound EW29Ch in subdomain α (left), subdomain β (centre) and subdomain γ (right).

Fig. S5. Plots of the order parameter (S2) and conformational exchange broadening factor (Rex) for the backbone of EW29Ch in the sugar-free state (black) and in the lactose-bound states at [EW29Ch]/[lactose] = 1/1 (blue), 1/8 (red) and 1/80 (purple).

Table S1. Lactose occupancy levels of the two sugar-binding sites of EW29Ch at each of the [EW29Ch]/[lactose] molar ratios.

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