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note
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50/[I])k}, where k is the Hill coefficient. For mechanism studies, data were analyzed using a nonlinear regression fit according to classical Michaelis-Menten kinetics models.
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33947584959
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note
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The reversibility of inhibition is easily determined by measuring the recovery of enzymatic activity after dialysis of the enzyme-inhibitor complex against assays. To test for reversibility, excessive compounds A4 and A17 were incubated with 100 nM enzyme. Then the mixture was dialyzed against the assay buffer and the activity of the enzyme was tested in different times. If the inhibitor was reversible, enzymatic activity could recover to some extent; and if the inhibitor was irreversible, enzymatic activity could not recover at any time. Figure 2 shows the activity of JSP-1 was recovered to some extent after dialysis, compared with the mixture without dialysis. It verified that compounds A4 and A17 were reversible JSP-1 inhibitors.
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