Mechanism of arsenite-mediated decreases in CYP3A23 in rat hepatocytes.
In primary cultures of rat hepatocytes, exposure to arsenite causes a major decrease in dexamethasone (DEX)-mediated induction of CYP3A23 hemoprotein, with a minor decrease in CYP3A23 mRNA. Here we show that addition of heme did not prevent the arsenite-mediated decreases in CYP3A23 protein, and arsenite did not decrease intracellular glutathione levels, indicating that heme and glutathione were not limiting for formation of holoCYP3A23. We also investigated whether arsenite decreases CYP3A23 protein by increasing CYP3A23 degradation by the calpain pathway. The calpain inhibitor, calpeptin, caused greater than a 90% inhibition of calpain-mediated proteolysis, but had no effect on DEX-mediated induction of CYP3A23 protein following 24h treatments. However, calpeptin enhanced the effect of arsenite to decrease induction of CYP3A23 protein. In addition, in short-term studies, calpeptin appeared to be a suicidal inhibitor of CYP3A-catalyzed enzyme activity. Our findings suggest that CYP3A23 protein is not degraded by calpain-mediated proteolysis, even in the presence of arsenite.
Veterans Administration Medical Center, White River Junction, VT, USA.
Trisha L
TL
Noreault
Judith M
JM
Jacobs
Ralph C
RC
Nichols
Heidi W
HW
Trask
Steven A
SA
Wrighton
Peter R
PR
Sinclair
Jacqueline F
JF
Sinclair
D000818:Animals; D018053:Arsenites / Q000008:administration & dosage*; D001189:Aryl Hydrocarbon Hydroxylases / Q000378:metabolism*; D002154:Calpain / Q000378:metabolism*; D002478:Cells, Cultured; D051544:Cytochrome P-450 CYP3A; D004151:Dipeptides / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005978:Glutathione / Q000378:metabolism*; D006418:Heme / Q000378:metabolism*; D022781:Hepatocytes / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D051381:Rats; D011916:Rats, Inbred F344; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology
D018053:Arsenites; D004151:Dipeptides; C071482:calpeptin; D006418:Heme; D001189:Aryl Hydrocarbon Hydroxylases; C465138:Cyp3a23-3a1 protein, rat; D051544:Cytochrome P-450 CYP3A; D002154:Calpain; D005978:Glutathione; C015001:arsenite