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For Oah assay, the reaction system contains 0.18 mM MnCl 2 and 0.040 M of purified Pcoah in 100 mM Tris-HCl buffer (pH = 8.0) with the final volume 0.5 mL

Ameen F, Alown F, Dawoud T, Sharaf A, Sakayanathan P, Alyahya S

1 H NMR (400 MHz, D 2 O) 7.78 (s, 2H), 7.01 (s, 2H), 4.71 4.60 (m, 2H), 4.44 4.03 (m, 3H), 3.80 3.74 (m, 4H), 3.32 2.88 (m, 10H), 1.85 1.11 (m, 18H) 13 .C NMR (151 MHz, D 2 O) 178.88, 172.79, 172.16, 172.10, 167.38, 167.21, 134.76, 134.56, 129.96, 129.66, 117.45, 54.20, 54.17, 53.68, 53.46, 53.08, 40.64, 40.59, 39.33, 39.28, 37.38, 30.79, 30.39, 30.37, 28.05, 27.65, 27.48, 27.40, 22.63, 22.18, 22.04

Importantly, 1a R -IN-8 with its reversible cyclohexenone-based warhead inhibited none of the 20 off-target kinases (Supplementary Table 3)