TABLE 1

Vmax values of wild-type hCE-2 and its variants toward irinotecan, p-nitrophenyl acetate, and 4-methylumberiferyl acetate Carboxylesterase activities toward the three substrates were determined as described under Materials and Methods. Concentrations used were 1 to 50 μM for irinotecan, 39 to 5000 μM for p-nitrophenyl acetate, and 62.5 to 1000 μM for 4-methylumbelliferyl acetate. Vmax values were determined by three independent experiments and were expressed as mean ± S.D. Data were expressed as pmol/mg microsomal protein/min for irinotecan, and as μmol/mg microsomal protein/min for p-nitrophenyl acetate and 4-methylumbelliferyl acetate.


Substrates

Wild-Type

R34W

V142M
Irinotecan 1.458 ± 0.0495 <0.2 <0.2
p-Nitrophenyl acetate 0.573 ± 0.0155 0.0513 ± 0.0073 0.0232 ± 0.0128
4-Methylumbelliferyl acetate
0.193 ± 0.0068
<0.03
<0.03