Skip to main content
. 2022 Aug 15;13:4785. doi: 10.1038/s41467-022-32436-4

Fig. 3. The S280F substitution causes changes in inhibitor potency varying with the active site substitution and type of inhibitor.

Fig. 3

a IC50 values (standard error of the mean, n = 3) with IDH1 variants (at 30 nM); n.i. = no inhibition observed. Conditions: 100 mM Tris, 10 mM MgCl2, 0.2 mM DTT, 0.005%v/v Tween 20, and 0.1 mg/mL BSA (pH 8.0). b Non-denaturing MS analyses. Dashed line: IDH1 dimer (z = 20, with 2 NADP(H) molecules bound), green shading corresponds to binding of one inhibitor molecule, mint shading corresponds to binding of 2 inhibitor molecules. Conditions: 20 µM IDH1 variant, 20 µM inhibitor, cone-voltage: 100 V. c KD determinations (standard errors of the mean) measured by by non-denaturing MS (20 µM enzyme, technical errors: n = 3 for the z = 19, 20, 21 charge states) and CPMG NMR (values in brackets, 10 µM enzyme, n = 2). Non-denaturing MS: ammonium citrate buffer (200 mM, pH 7.5); CPMG NMR: 50 mM Tris-d11, 100 mM NaCl, 10 mM MgCl2, and 10% D2O, pH 7.5.