Metabolite | Retention Time | M +H+ | Rate of Metabolite Formation | |||
---|---|---|---|---|---|---|
NADPH | NADPH + UDPGA | NADPH + ala | NADPH + UDPGA + ala | |||
min | pmol/min-mg protein | |||||
M1 | 12.0 | 207 | 475 ± 9 | 222 ± 7 | 477 ± 8 | 87 ± 1 |
M2 | 8.9 | 383 | N.D. | 345 ± 5 | N.D. | 500 ± 2 |
M3 | 7.9 | 163 | 582 ± 2 | 261 ± 20 | 608 ± 11 | 100 ± 1 |
M4 | 4.0 | 339 | N.D. | 292 ± 5 | N.D. | 502 ± 4 |
Metabolites M3 and M4 were identified as 7-HC and 7-HC glucuronide, respectively. Formation rates for metabolites M1 and M2 were estimated using the standard curves for 7-HC and 7-HC glucuronide, respectively. Some of these data have been published previously (Fisher et al., 2000).
ala, alamethicin (added as described under Materials and Methods); N.D., not detected.