Ring-andN-hydroxylation of 2-acetamidofluorene by Rat Liver Reconstituted Cytochrome P-450 Enzyme System

Overview

Journal Biochem J

Specialty Biochemistry

Date 1975 Sep 1

PMID 813632

Citations 6

Authors

P D LOTLIKAR

K Zaleski

Affiliations

Soon will be listed here.

Abstract

The N- and ring-hydroxylation of 2-acetamidofluorene were studied with a reconstituted cytochrome P-450 enzyme from microsomal fractions of liver from both control and 3-methylcholanthrene-pretreated rats. Proteinase treatment and Triton X-100 solubilization were two important steps for partial purification of the cytochrome P-450 fraction. Both cytochrome P-450 and NADPH-cytochrome c reductase fractions were required for optimum N- and ring-hydroxylation activity. Hydroxylation activity was determined by the source of cytochrome P-450 fraction; cytochrome P-450 fraction from pretreated animals was severalfold more active than the fraction from controls. Formation of N-hydroxylated metabolites with reconstituted systems from both control and pretreated animals was greater than that with their respective whole microsomal fractions.

Citing Articles

Pathobiologic and metabolic aspects of mammary gland tumorigenesis by N-substituted aryl compounds.

Ritter C, Ryzewski C Environ Health Perspect. 1983; 49:175-83.

PMID: 6339224 PMC: 1569135. DOI: 10.1289/ehp.8349175.

Effect of detergents on the N-and ring-hydroxylation of 2-acetamidofluorene by hamster liver microsomal preparations.

LOTLIKAR P, Dwyer E Biochem J. 1976; 160(3):821-4.

PMID: 1016257 PMC: 1164302. DOI: 10.1042/bj1600821.

Phospholipid requirement for dimethylnitrosamine demethylation by hamster hepatic microsomal cytochrome P-450 enzyme system.

LOTLIKAR P, Baldy Jr W, Nyce J, Dwyer E Biochem J. 1976; 160(2):401-4.

PMID: 1008863 PMC: 1164247. DOI: 10.1042/bj1600401.

Dimethylnitrosamine demethylation by reconstituted liver microsomal cytochrome P-450 enzyme system.

LOTLIKAR P, Baldy Jr W, Dwyer E Biochem J. 1975; 152(3):705-8.

PMID: 819003 PMC: 1172528. DOI: 10.1042/bj1520705.

Phospholipid requirement for 2-acetamidofluorene N-and ring-hydroxylation by hamster liver microsomal cytochrome P-450 enzyme system.

LOTLIKAR P, Dwyer E, Baldy Jr W, Nyce J Biochem J. 1977; 168(3):571-4.

PMID: 606254 PMC: 1183807. DOI: 10.1042/bj1680571.

References

Lu A, West S . Reconstituted liver microsomal enzyme system that hydroxylates drugs, other foreign compounds, and endogenous substrates. 3. Properties of the reconstituted 3,4-benzypyrene hydroxylase system. Mol Pharmacol. 1972; 8(5):490-500. View

Lu A, Levin W, West S, Jacobson M, Ryan D, Kuntzman R . The role of cytochrome P-450 and P-448 in drug and steroid hydroxylations. Ann N Y Acad Sci. 1973; 212:156-74. DOI: 10.1111/j.1749-6632.1973.tb47594.x. View

Strobel H, Lu A, Heidema J, Coon M . Phosphatidylcholine requirement in the enzymatic reduction of hemoprotein P-450 and in fatty acid, hydrocarbon, and drug hydroxylation. J Biol Chem. 1970; 245(18):4851-4. View

Lu A, West S, Vore M, Ryan D, Levin W . Role of cytochrome b5 in hydroxylation by a reconstituted cytochrome P-450-containing system. J Biol Chem. 1974; 249(21):6701-9. View

LOTLIKAR P, Luha L, Zaleski K . Reconstituted hamster liver microsomal enzyme system for N-hydroxylation of the carcinogen 2-acetylaminofluorene. Biochem Biophys Res Commun. 1974; 59(4):1349-55. DOI: 10.1016/0006-291x(74)90462-8. View

LOTLIKAR P . Effects of 3-methylcholanthrene pretreatment on microsomal hydroxylation of 2-acetamidofluorene by various rat hepatomas. Biochem J. 1970; 118(3):513-8. PMC: 1179220. DOI: 10.1042/bj1180513. View

WEISBURGER J, WEISBURGER E . Biochemical formation and pharmacological, toxicological, and pathological properties of hydroxylamines and hydroxamic acids. Pharmacol Rev. 1973; 25(1):1-66. View

LOTLIKAR P, Zaleski K . Inhibitory effect of carbon monoxide on the N- and ring-hydroxylation of 2-acetamidofluorene by hamster hepatic microsomal preparations. Biochem J. 1974; 144(2):427-30. PMC: 1168511. DOI: 10.1042/bj1440427. View

Miller J . Carcinogenesis by chemicals: an overview--G. H. A. Clowes memorial lecture. Cancer Res. 1970; 30(3):559-76. View

10.

LOTLIKAR P, Enomoto M, Miller J, Miller E . Species variations in the N- and ring-hydroxylation of 2-acetylaminofluorene and effects of 3-methylcholanthrene pretreatment. Proc Soc Exp Biol Med. 1967; 125(2):341-6. DOI: 10.3181/00379727-125-32086. View

11.

Alvares A, Schilling G, Levin W, Kuntzman R . Studies on the induction of CO-binding pigments in liver microsomes by phenobarbital and 3-methylcholanthrene. Biochem Biophys Res Commun. 1967; 29(4):521-6. DOI: 10.1016/0006-291x(67)90515-3. View

12.

Miller J, Cramer J, Miller E . The N- and ringhydroxylation of 2-acetylaminofluorene during carcinogenesis in the rat. Cancer Res. 1960; 20:950-62. View

13.

Omura T, Sato R . THE CARBON MONOXIDE-BINDING PIGMENT OF LIVER MICROSOMES. I. EVIDENCE FOR ITS HEMOPROTEIN NATURE. J Biol Chem. 1964; 239:2370-8. View

14.

Miller E, Miller J, Brown R, MacDonald J . On the protective action of certain polycyclic aromatic hydrocarbons against carcinogenesis by aminoazo dyes and 2-acetylaminofluorene. Cancer Res. 1958; 18(4):469-77. View

15.

LOWRY O, ROSEBROUGH N, FARR A, RANDALL R . Protein measurement with the Folin phenol reagent. J Biol Chem. 1951; 193(1):265-75. View

16.

Lu A, Levin W . The resolution and reconstitution of the liver microsomal hydroxylation system. Biochim Biophys Acta. 1974; 344(2):205-40. DOI: 10.1016/0304-4157(74)90004-5. View

17.

Thorgeirsson S, JOLLOW D, SASAME H, Green I, Mitchell J . The role of cytochrome P-450 in N-hydroxylation of 2-acetylaminofluorene. Mol Pharmacol. 1973; 9(3):398-404. View

18.

Masters B, Baron J, TAYLOR W, Isaacson E, LOSPALLUTO J . Immunochemical studies on electron transport chains involving cytochrome P-450. I. Effects of antibodies to pig liver microsomal reduced triphosphopyridine nucleotide-cytochrome c reductase and the non-heme iron protein from bovine adrenocortical.... J Biol Chem. 1971; 246(13):4143-50. View

19.

van der Hoeven T, Coon M . Preparation and properties of partially purified cytochrome P-450 and reduced nicotinamide adenine dinucleotide phosphate-cytochrome P-450 reductase from rabbit liver microsomes. J Biol Chem. 1974; 249(19):6302-10. View

20.

Lu A, Junk K, Coon M . Resolution of the cytochrome P-450-containing omega-hydroxylation system of liver microsomes into three components. J Biol Chem. 1969; 244(13):3714-21. View