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Fluorescence-based inhibition assay for L. donovani CYP51 (A) and inhibition by 24c (B). The fluorogenic substrate BFC (50 μM) was incubated with purified CYP51-32c (60 nM) in the presence of cofactor CuOOH (0.1 mM) or hydrogen peroxide (HOOH; 25 mM) at 37°C. Control incubations did not contain either the cofactor (but with CYP) or the CYP protein (but with CuOOH or HOOH). IC50 value (mean ± SE of triplicate determinations) is shown for 24c.
