Effect of CYP2A13-catalyzed nicotine metabolism on coumarin 7-hydroxylation, reduced CO spectrum, and HPLC-detected hemea
Percent of Controlb | |||||
|---|---|---|---|---|---|
| Activity Remaining | Reduced CO Spectrum | Heme Remaining | |||
| Before spin column/dialysis (n = 4) | 40 ± 3c | 69 ± 5c | 64 ± 5c | ||
| After spin column (n = 2 or 3) | 36 ± 6c | 54 ± 26 | 71 | ||
| After dialysis (n = 2 or 3) | 35 ± 5c | 45 | 59 | ||
↵ a Reconstituted CYP2A13 was incubated with 25 μM nicotine for 16 min at 30°C prior to assaying for activity in a secondary reaction and reduced CO spectra and heme in the primary incubation as described under Materials and Methods
↵ b Values are means ± S.D. from two to four separate experiments. When only two values were averaged, they did not differ by greater than 10%. Control samples contained no NADPH
↵ c Value less than control value, P < 0.001