Table 1.
Influx kinetics for pemetrexed and methotrexate for wt-PCFT and the W299S mutant
wt-PCFT | W299S | W299S/wt-PCFT | |
---|---|---|---|
Pemetrexed Kt, μM | 0.99 ± 0.23 | 0.28 ± 0.01 | 0.28 |
Pemetrexed Vmax, pmol/mg protein | 384 ± 83 | 58.3 ± 8.7 | 0.15 |
Pemetrexed Ki, μM | 0.94 ± 0.07 | 0.16 ± 0.02 | 0.17 |
MTX Ki, μM | 2.36 ± 0.06 | 1.05 ± 0.03 | 0.44 |
Values are the mean ± SE from 3 independent experiments. Influx Kt, Ki and Vmax were computed from the Michaelis-Menton equation. All differences between wild-type proton-coupled folate transporter (PCFT) (wt-PCFT) and the W299S mutant are significant to P ≤ 0.01. The difference between the fold decrease in the influx Ki for the W299S mutant relative to wt-PCFT for both [3′,5′,7-3H]methotrexate (MTX) and pemetrexed is significant (P < 0.03). The difference between MTX Ki and pemetrexed Ki for wt-PCFT as well as the difference between the W299S mutants is significant (P < 0.003).