FIGURE 2.

ZINC12899676 inhibits the activity of PEDV NTPase. (A) Purification of PEDV nsp13. “E” represents empty vector, “L” represents whole bacteria after induction, “S” represents the supernatant after ultrasonication, “B” represents the liquid collected after washing using binding buffer, “W” represents the liquid collected after washing using washing buffer, and numbers (1–8) represent the liquid collected after washing using elution buffer. (B) Purified nsp13 recombinant proteins (0, 0.1, 0.2, or 0.5 μM) were incubated with ATP in the reaction buffer at 37°C for 20 min, and then, the Kinase-Glo reagent mix was added, and the ATPase activity was measured. (C) PEDV nsp13 (0.2 μM) was reacted with ATP at 37°C for 5, 10, 20, 30, or 40 min, and then, the ATPase activity was measured. (D) Compounds (25 μM) or DMSO were added to the reaction mixture 10 min ahead of ATP, and the ATPase activity was measured. (E) Different concentrations of ZINC12899676 were added to the reaction mixture 10 min ahead of ATP, and the ATPase activity was measured. (F) 2D structure of ZINC12899676.