| Assay Method Information | |
| | MenE-MenB Coupled Assay |
| Description: | Enzyme inhibition studies were performed in 20 mM NaHPO4 buffer (pH 7.4) containing 150 mM NaCl and 1 mM MgCl2 using a MenE-MenB coupled assay in whichMenE is rate-limiting. IC50 values were determined in reaction mixtures containing OSB (60 μM), ATP (240 μM), CoA (240 μM), mtMenB (2.5 μM), and varying inhibitor concentrations (5-250 μM). Reactions were initiated by addition of ecMenE (50 nM), and the production of DHNACoA was monitored at 392 nm (ε392 = 4000 M-1 cm-1). |
| Affinity data for this assay | |
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