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| | Inhibition Kinetics Assay |
| Description: | Binding of TLM18 to wild-type and mutant KasA was quantified by monitoring changes in the intrinsic tryptophan fluorescence of the enzyme at wavelengths of 280 nm for excitation and 337 nm for emission, as described previously [Kapilashrami et al., J. Biol. Chem., 288:6045-6052]. Briefly, the data were collected using a Quanta Master fluorometer (Photon Technology International). Inhibitors were dissolved in DMSO and titrated into 1 μM solutions of KasA and C171Q KasA in buffer (50 mM Tris-HCl, 150 mM NaCl, 1 mM dithiothreitol, pH 8.5). |
| Affinity data for this assay | |
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