Kinetics of acetyl coenzyme A:arylamine N-acetyltransferase from rapid and slow acetylator frog tissues.
N-acetyltransferase (NAT) activity was determined in 100 frog (Rana tigrina) livers using 2-aminofluorene and p-aminobenzoic acid as substrates. Overall, the liver NAT activity of the 50 females was higher than the liver NAT activity of the 50 males. The activities (mean +/- SD) of NAT from the bladder, blood, colon, and liver of males was 0.30 +/- 0.11, 0.05 +/- 0.03, 0.09 +/- 0.05, and 0.93 +/- 0.56 nmol/min/mg protein for the acetylation of aminofluorene and 0.29 +/- 0.06, 0.36 +/- 0.04, 0.26 +/- 0.02, and 0.32 +/- 0.14 nmol/min/mg protein for the acetylation of p-aminobenzoic acid. In the bladder, blood, colon, and liver from female frogs, the activities obtained were 1.00 +/- 0.41, 0.52 +/- 0.07, 0.08 +/- 0.05, and 1.27 +/- 0.49 nmol/min/mg protein for aminofluorene and 0.34 +/- 0.12, 0.36 +/- 0.04, 0.34 +/- 0.07, and 0.48 +/- 0.21 nmol/min/mg protein for p-aminobenzoic acid. Kinetic constants for arylamine NAT activity in the blood, liver, bladder, and colon from frogs with rapid, intermediate, and slow acetylator activities were determined. KM and Vmax values for aminofluorene were 2- to 6-fold higher for liver than for the other tissues. KM and Vmax values for p-aminobenzoic acid showed a smaller variation among the tissues examined, with values obtained for the liver and bladder being somewhat higher than the values for the blood and colon. An apparent KM difference for aminofluorene was found in the liver from frogs with high and low acetylator activity. Based on the aminofluorene NAT activity of liver, there seems to be a polymorphism in NAT activity with 4 rapid, 21 intermediate, and 75 slow acetylators among the 100 frogs assayed. Distribution of acetylator phenotypes was similar among the 50 males and 50 females in this study. This is the first demonstration of acetyl coenzyme A:arylamine NAT activity in an amphibian and could lead to the development of a frog model for monitoring the effect of pollution of wetland environments on native species.
Ho
C C
CC
Department of Medicine, China Medical College, Taiwan, Republic of China.
Lin
T H
TH
Lai
Y S
YS
Chung
J G
JG
Levy
G N
GN
Weber
W W
WW
D000105:Acetyl Coenzyme A / Q000737:chemistry*; D000107:Acetylation; D000818:Animals; D001191:Arylamine N-Acetyltransferase / Q000097:blood / Q000737:chemistry / Q000378:metabolism*; D003106:Colon / Q000201:enzymology*; D005260:Female; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008297:Male; D010641:Phenotype; D011898:Ranidae; D001743:Urinary Bladder / Q000201:enzymology*
D000105:Acetyl Coenzyme A; D001191:Arylamine N-Acetyltransferase