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. Author manuscript; available in PMC: 2021 May 23.
Published in final edited form as: Nat Metab. 2020 Nov 23;2(12):1413–1426. doi: 10.1038/s42255-020-00313-3

Extended Data Fig. 2. HEX is a substrate-competitive inhibitor with preference for ENO2.

Extended Data Fig. 2

a, b. Enolase activity (1/v, y-axis) was measured spectrophotometrically in vitro using an NADH-coupled assay1,2, with ENO1 and ENO2 activity plotted as a function of substrate concentration (x-axis, 1/S: 2-phosphoglycerate in mM; HEX in nM). Data were plotted as Lineweaver-Burke with x-axis showing inverse of substrate versus inverse activity (1/V). Each dot represents on independent biochemical rate determination. Slopes (Km-apparent/Vmax) were re-plotted as a function of inhibitor concentration in Figure 1c. c. Enolase activity in lysates from ENO1 and ENO2 overexpressing D423 cells, measured as a function of HEX concentration; 0.5 mM 2-PG substrate concentration.