Purification and characterization of cytochrome P-450 isozymes from phenobarbital-induced adult hen liver.
Gupta-RP; Lapadula-DM; Abou-Donia-MB
Comp Biochem Physiol C Comp Pharmacol Toxicol 1990 Jan; 96(1):163-176
Cytochrome-P-450 isozymes from the livers of adult hens treated with phenobarbital (50066) were purified and characterized. Leghorn-hens were injected intraperitoneally with phenobarbital at 20mg/kg daily for 4 days; 24 hours after the last dose, they were killed, and liver microsomes were prepared. P-450 isozymes were separated by column chromatography. The isozymes were identified as P-450 PB-A and PB-B. Two highly specific P-450 antibodies were raised for both the isozymes, and these produced a single band with microsomes by immunoblotting. Each of the two purified P-450 isozymes had the same molecular weight, 54,000. However, each was easily distinguished using chromatographic behavior, spectral analysis, peptide mapping, tryptic peptide mapping, immunochemical properties, and amino terminal sequencing. There was no cross reactivity between the antibodies raised against P-450 PB-A and PB-B, and microsomal P450s of rat, mice, cat, or catfish species as determined by immunoblotting.
NIOSH-Publication; NIOSH-Grant; Neurotoxic-effects; Laboratory-animals; Liver-enzymes; Microsomal-enzymes; Enzyme-activity; Oxidative-enzymes; Immunochemistry
Pharmacology Duke University Department of Pharmacology Durham, N C 27710
Neurotoxic Disorders; Neurotoxic-effects
Comparative Biochemistry and Physiology, Part C: Comparative Pharmacology and Toxicology
Duke University, Durham, North Carolina