Kinetic modeling of DNA adduct yields to obtain kinetic parameters. Yields of AFB diol obtained with 18 μM AFB
1 exo-8,9-epoxide and varying concentrations of DNA were fit to the model:
where A = AFB
1 exo-8,9-epoxide, D = AFB diol, B = DNA, C = DNA–AFB adduct, and
k0 was determined to be 0.60 s
−1 (
4).
Kd and
kcat were estimated by reiterative fitting with a
KINSIM program (
11) to give values of C that are consistent with the experimentally determined values (of C and D) found with varying concentration of A and B (
12). The solid line is that for
Kd = 0.43 mg ml
−1 (1.4 mM monomer equivalents) and
kcat = 35 s
−1.