Full kinetic schemes for ecDHFR and hDHFR. hDHFR rates
are shown
in bold, and ecDHFR rates are shown in parentheses. Black arrows indicate
the major catalytic cycle for both ecDHFR and hDHFR. The minor “upper”
cycle, shown by blue arrows, is also used by hDHFR because the release
of THF from the product complex is much faster than for ecDHFR. Approximately
35% of the product is channeled through the upper branch by human
DHFR. Gray arrows show kinetics involving the apoprotein, E, which
do not constitute a significant part of the enzymatic cycle for either
protein under cellular conditions. ecDHFR kinetic parameters (25 °C,
pH 7) were taken from ref (39), and hDHFR parameters (20 °C, pH 7.65) were taken
from ref (14). First-
and second-order rates are given in units of s–1 and μM–1 s–1, respectively.