Induction of rat hepatic N-nitrosodimethylamine demethylase by acetone is due to protein stabilization.
The N-nitrosodimethylamine demethylase (P450I-IE1) is induced severalfold in liver by giving rats ethanol, acetone, pyrazole, and other related small molecular weight compounds. This induction is not the result of an increase in IIE1 mRNA, but could be due to either an increase in translation rate or a decrease in protein degradation. To determine the mechanism of induction, we measured IIE1 synthesis and degradation rates in untreated and acetone-treated rats. This was accomplished by immunopurification of radiolabeled IIE1 protein using a specific monoclonal antibody subsequent to in vivo labeling of total cellular protein with either NaH14CO3 or [3H]leucine. We found that in rats fed acetone, the rate of IIE1 synthesis was not changed; however, IIE1 degradation was markedly altered. In untreated rats, IIE1 protein was degraded via a biphasic pathway consisting of both a rapid and slow component with approximate half-lives of 7 and 37 h, respectively. However, in acetone-treated rats, only a monophasic curve with a half-life of 37 h was observed. The abolition of the rapid degradation component of the IIE1 turnover cycle indicates that induction of IIE1 by acetone is primarily due to specific stabilization of IIE1 protein. Since acetone is also metabolized by IIE1, we believe that this may be a substrate-induced enzyme stabilization.
Song
B J
BJ
Laboratory of Metabolism and Molecular Biology, National Institute on Alcohol Abuse and Alcoholism, Rockville, Maryland 20852.
Veech
R L
RL
Park
S S
SS
Gelboin
H V
HV
Gonzalez
F J
FJ
D000096:Acetone / Q000494:pharmacology*; D000818:Animals; D001345:Autoradiography; D019392:Cytochrome P-450 CYP2E1; D004591:Electrophoresis, Polyacrylamide Gel; D004790:Enzyme Induction / Q000187:drug effects; D007700:Kinetics; D007930:Leucine / Q000378:metabolism; D008297:Male; D008862:Microsomes, Liver / Q000201:enzymology*; D010089:Oxidoreductases, N-Demethylating / Q000096:biosynthesis* / Q000302:isolation & purification / Q000378:metabolism; D014176:Protein Biosynthesis / Q000187:drug effects; D011720:Pyrazoles / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains
D011720:Pyrazoles; D000096:Acetone; C031280:pyrazole; D019392:Cytochrome P-450 CYP2E1; D010089:Oxidoreductases, N-Demethylating; D007930:Leucine