| Assay Method Information | |
| | Steady-State Kinetics |
| Description: | Initial rates of amine oxidation were routinely measured by monitoring oxygen consumption in 0.1 M Hepes (pH 8.0) and 0.1 M NaCl at 25 °C using an oxygen electrode (model 5300A, Yellow Springs Instruments, Yellow Springs, OH). With (R,S)-6-hydroxynicotine and (S)-6-hydroxynornicotine, the concentration of enzyme was typically 0.1 μM; this was increased to 4 μM for (S)-nicotine and (R,S)-4-(1-methyl pyrrolidine-2-yl)phenol. The concentration of oxygen was varied by bubbling the desired concentration of oxygen (62 μM to 1.25 mM) into the oxygen electrode cell for ∼10 min. |
| Affinity data for this assay | |
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