Extended Data Figure 6: Representative ITC raw data and binding isotherms for MD2 interacting with mutant MraYAA.
From: Structural insights into inhibition of lipid I production in bacterial cell wall synthesis
All titrations were performed in triplicate (technical replicates); see source data for all titrations. Representative data are shown. For MraYAA mutants Lys70Ala, Thr75Ala, Asp193Asn, Asn255Ala, Phe262Trp and His325Ala, 210 μM MD2 was titrated into 30 μM enzyme. For MraYAA Gln305Ala, 315–430 μM MD2 was titrated into 25–27 μM enzyme. For MraYAA Phe262Ala, 80–110 μM MD2 was titrated into 7–10.5 μM enzyme. Mean thermodynamic parameters for triplicate titrations are shown in the Extended Data Table 2. Mean Kd values for each triplicate are as follows: 63.9 ± 4.7 nM for Lys70Ala; 27.4 ± 1.5 nM for Thr75Ala; Kd not determined for Asp193Asn; 29.7 ± 0.8 nM for Asn255Ala; 228 ± 4 nM for Phe262Ala; 68.4 ± 0.9 nM for Phe262Trp; 117 ± 10 nM for Gln305Ala; 24.4 ± 0.4 nM for His325Ala.
