ON THE MECHANISM OF DRUG HYDROXYLATION IN RAT LIVER MICROSOMES

The TPNH- and O2-dependent drug hydroxylation system of liver microsomes has been studied using normal rats and rats in which the drug-hydroxylating activity has been enhanced by repeated injections of phenobarbital. The oxidative demethylation of aminopyrine is employed as an assay. Optimal conditions for the assay with regard to the concentrations of TPNH and aminopyrine are established. TPN inhibits the reaction in a competitive manner, similarly to its effect on the microsomal TPNH-cytochrome c reductase. Drug hydroxylation, but not the "TPNH oxidase," TPNH-cytochrome c, -2,6-dichlorophenolindophenol, or -neotetrazolium reductase reaction, or the TPNH-dependent lipid peroxidation, is blocked by carbon monoxide. Microsomes from phenobarbital-treated rats exhibit increased activities of the various TPNH-linked reductase reactions, parallel to the increased drug hydroxylation activity, whereas the "TPNH oxidase" activity does not change appreciably. Measurements with microsomes from drug-treated animals reveal a 1:1:1 stoichiometry of aminopyrine-dependent oxygen uptake, TPNH oxidation, and formaldehyde formation. Attempts to solubilize the drug-hydroxylating enzyme system are also presented. It is concluded that the drug-hydroxylating enzyme system involves the microsomal TPNH-cytochrome c reductase and CO-binding pigment, and a hypothetic reaction scheme accounting for the data presented is proposed.

• Publication year

  1965

• Venue

  Journal of Cell Biology

• Publication date

  1965-09-01

• Fields of study

  Biology, Medicine, Chemistry

• Identifiers
  DOI 10.1083/JCB.26.3.713PMID 19866674PMCID 2106795
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

• FURTHER STUDIES ON THE INDUCTION OF THE DRUG-HYDROXYLATING ENZYME SYSTEM OF LIVER MICROSOMES

  1965cited by this paper
• PHENOBARBITAL-INDUCED SYNTHESIS OF THE MICROSOMAL DRUG-METABOLIZING ENZYME SYSTEM AND ITS RELATIONSHIP TO THE PROLIFERATION OF ENDOPLASMIC MEMBRANES

  1965cited by this paper
• Evidence for the involvement of iron in the ADP-activated peroxidation of lipids in microsomes and mitochondria.

  1964cited by this paper
• THE CARBON MONOXIDE-BINDING PIGMENT OF LIVER MICROSOMES. I. EVIDENCE FOR ITS HEMOPROTEIN NATURE.

  1964cited by this paper
• Ciba Foundation Symposium - Cellular Injury

  1964cited by this paper
• THE CARBON MONOXIDE-BINDING PIGMENT OF LIVER MICROSOMES. II. SOLUBILIZATION, PURIFICATION, AND PROPERTIES.

  1964cited by this paper
• Oxidases and Related Oxidation-Reduction Systems

  1964cited by this paper
• Inhibition of the TPNH-linked lipid peroxidation of liver microsomes by drugs undergoing oxidative demethylation.

  1964cited by this paper
• METABOLISM OF DRUGS AND TOXIC SUBSTANCES.

  1964cited by this paper
• ENERGY-DEPENDENT REDUCTION OF TRIPHOSPHOPYRIDINE NUCLEOTIDE BY REDUCED DIPHOSPHOPYRIDINE NUCLEOTIDE, COUPLED TO THE ENERGY-TRANSFER SYTEM OF THE RESPIRATORY CHAIN.

  1963cited by this paper
• THE LIGHT REVERSIBLE CARBON MONOXIDE INHIBITION OF THE STEROID C21-HYDROXYLASE SYSTEM OF THE ADRENAL CORTEX.

  1963cited by this paper
• ADP-ACTIVATED LIPID PEROXIDATION COUPLED TO THE TPNH OXIDASE SYSTEM OF MICROSOMES.

  1963cited by this paper
• Cofactor requirements of the O-demethylating liver microsomal enzyme system.

  1963cited by this paper
• [ON THE KINETICS OF MICROSOMAL NADPH OXIDATION].

  1963cited by this paper
• Ciba Foundation Symposium - Enzymes and Drug Action

  1962cited by this paper
• ENZYME-STRUCTURE RELATIONSHIPS IN THE ENDOPLASMIC RETICULUM OF RAT LIVER

  1962cited by this paper
• DT diaphorase. I. Purification from the soluble fraction of rat-liver cytoplasm, and properties.

  1962cited by this paper
• Hepatic triphosphopyridine nucleotide-cytochrome c reductase: isolation, characterization, and kinetic studies.

  1962cited by this paper
• Microsomal triphosphopyridine nucleotide-cytochrome c reductase of liver.

  1962cited by this paper
• Enzymatic hydroxylation of aromatic compounds.

  1961cited by this paper
• The metabolism of drugs by subfractions of hepatic microsomes.

  1961cited by this paper
• Solubilization of aromatic hydroxylase system of liver microsomes and requirement of lipid-like factor.

  1960cited by this paper
• A microsomal TPNH-neotetrazolium diaphorase.

  1959cited by this paper
• Studies of liver glucose 6-phosphatase. III. Solubilization and properties of the enzyme from normal and diabetic rats.

  1959cited by this paper
• Pigments of rat liver microsomes.

  1958cited by this paper
• The metabolism of methylated aminoazo dyes. VI. Intracellular distribution and properties of the demethylase system.

  1957cited by this paper
• The oxidation of drugs by liver microsomes: on the role of TPNH and oxygen.

  1957cited by this paper
• Pyridine nucleotide transhydrogenase. III. Animal tissue transhydrogenases.

  1953cited by this paper
• Protein measurement with the Folin phenol reagent.

  1951cited by this paper
• The reaction between thiobarbituric acid and the oxidation products of certain lipides.

  1948cited by this paper
• The colorimetric determination of phosphorus.

  1932cited by this paper
• Cancer Research

  1902cited by this paper
• Federation of European Biochemical Societies

  year unknowncited by this paper

• In memoriam: Sten Orrenius (1937-2020).

  2020cites this paper
• Role of Cell Death in Toxicology: Does It Matter How Cells Die?

  2019cites this paper
• In vivo characterization of TMX1: TMX1 preferentially acts upon transmembrane polypeptides

  2016cites this paper
• EFFECT OF CYCLOHEXIMIDE TREATMENT ON THE MIXED-FUNCTION OXIDASE SYSTEM OF RAT LIVER

  2012cites this paper
• Early work on apoptosis, an interview with Sten Orrenius

  2006cites this paper
• Über die Rolle von Sulfhydrylgruppen bei der mikrosomalen Arzneimitteldemethylierung

  2004cites this paper
• Induction of liver microsomal monooxygenases in experimental cholestasis

  2004cites this paper
• Die oxydative N-demethylierung von N-monomethyl-p-nitranilin

  2004cites this paper
• Über den Einfluß induzierender Substanzen auf Fremdstoff-Oxydasen und andere Redoxenzyme der Leber

  2004cites this paper
• Age-related effects of heat stress on protective enzymes for peroxides and microsomal monooxygenase in rat liver.

  1997cites this paper
• Effects of flavonoids on cytochrome P‐450 from rat liver microsomes: Inhibition of enzyme activities and protection against peroxidative damage

  1995cites this paper
• Effect of hyperthermia on glutathione peroxidase and lipid peroxidative damage in liver

  1994cites this paper
• Microbial models of mammalian metabolism: involvement of cytochrome P450 in the N-demethylation of N-methylcarbazole by Cunninghamella echinulata.

  1993cites this paper
• Generation of free radicals during the reductive metabolism of the nitroaromatic compound, nilutamide.

  1991cites this paper
• Mutagenicity of 3‐chlorodibenzofuran and its metabolic activation

  1991cites this paper
• Two sites of azo reduction in the monooxygenase system.

  1988cites this paper
• Xenobiotic metabolism in higher plants involvement of microsomal cytochrome P-450 in aminopyrine N-demethylation

  1988cites this paper
• The one-electron reduction of uroporphyrin I by rat hepatic microsomes.

  1987cites this paper
• [An organic peroxide, methyl ethyl ketone peroxide, damage to cytochrome P-450 peroxidase and microsomal enzymes activity].

  1986cites this paper
• A quantitative histochemical study ferrihemoprotein reductase activity of NADPH

  1986cites this paper
• A quantitative histochemical study of NADPH-ferrihemoprotein reductase activity

  1986cites this paper
• Alteration of hepatocellular structure and function by thallium chloride: ultrastructural, morphometric, and biochemical studies.

  1986cites this paper
• Sex-dependent activities of quinone reductases in rabbits indicate higher risk of bladder cancer in the male.

  1986cites this paper
• Evidence for a predominantly NADH-dependent O-dealkylating system in rat hepatic microsomes.

  1985cites this paper
• Effect of dietary vitamin E on methyl ethyl ketone peroxide damage to microsomal cytochrome P-450 peroxidase.

  1985cites this paper
• Methyl ethyl ketone peroxide damage to cytochrome P-450 peroxidase activities.

  1985cites this paper
• Activities of several phase I and phase II xenobiotic biotransformation enzymes in cultured hepatocytes from male and female rats.

  1985cites this paper
• Biliary cirrhosis and tumors induced by chronic administration of thiobenzamide to rats

  1984cites this paper
• Studies on the mechanisms of thallium-mediated inhibition of hepatic mixed function oxidase activity. Correlation with inhibition of NADPH-cytochrome c (P-450) reductase.

  1984cites this paper
• On the stoichiometry of the oxidase and monooxygenase reactions catalyzed by liver microsomal cytochrome P-450. Products of oxygen reduction.

  1984cites this paper
• Effect of Phototherapy on Hepatic Microsomal Drug Metabolism in Heterozygous and Homozygous Gunn Rats

  1984cites this paper
• Role of the microsomal FAD-containing monooxygenase in the liver toxicity of thioacetamide S-oxide.

  1984cites this paper
• Microsomal monooxygenase system in frog livers.

  1984cites this paper
• Alteration of hepatic microsomal structure and function by indium chloride. Ultrastructural, morphometric, and biochemical studies.

  1983cites this paper
• Generation of free radical metabolites and superoxide anion by the calcium indicators arsenazo III, antipyrylazo III, and murexide in rat liver microsomes.

  1983cites this paper
• Methimazole-induced modulation of thiobenzamide bioactivation and toxicity.

  1983cites this paper
• Detoxification of the phytoalexin pisatin by a fungal cytochrome P-450.

  1983cites this paper
• Drug-oxidizing mono-oxygenase system in liver microsomes of goldfish (Carassius auratus).

  1983cites this paper
• Endotoxin and low protein diet induced depression of the rat hepatic drug metabolism.

  1983cites this paper
• Influence of perinatal nutrition on hepatic drug metabolism in the adult rat.

  1983cites this paper
• Properties of NADH-dependent estradiol 2-hydroxylase in rat liver.

  1982cites this paper
• Spin-trapping studies on the free-radical products formed by metabolic activation of carbon tetrachloride in rat liver microsomal fractions isolated hepatocytes and in vivo in the rat.

  1982cites this paper
• Immobilization of liver microsomal cytochrome P‐450

  1982cites this paper
• Litcrosomal Nadph Oxidase and Hybrid Electrodes

  1982cites this paper
• Fibrin membrane endowed with biological function

  1982cites this paper
• Effects of endotoxin upon rat hepatic microsomal drug metabolism in vivo and in vitro.

  1982cites this paper
• Different behaviors of benznidazole as free radical generator with mammalian and Trypanosoma cruzi microsomal preparations.

  1982cites this paper
• Generation of free radicals induced by nifurtimox in mammalian tissues.

  1981cites this paper
• Nitrofuran enhancement of microsomal electron transport, superoxide anion production and lipid peroxidation.

  1981cites this paper
• Control of delta-aminolaevulinate synthase and haem oxygenase in chronic-iron-overloaded rats.

  1981cites this paper
• The presence of a microsomal UDP‐glucuronyl transferase for bilirubin in homozygous jaundiced gunn rats and in the crigler‐najjar syndrome

  1981cites this paper
• Stoichiometry of aminopyrine demethylation with and without NADH synergism.

  1981cites this paper
• Hepatic mixed-function oxidase of the mallard duck (Anas platyrhynchos).

  1981cites this paper
• Hepatic aminopyrine N-demethylase system: interaction of aminopyrine with microsomal cytochrome P-450.

  1981cites this paper
• Secretory Organelles: I The Endomembrane System

  1981cites this paper
• Liver growth after partial hepatectomy: influence of phenobarbital administration.

  1981cites this paper
• Alterations in the latency of hepatic microsomal hexose-6-phosphate dehydrogenase under various in vivo and in vitro conditions.

  1981cites this paper
• Decrease of the activity of the mixed function oxidase system in regenerating rat liver: an alternative explanation.

  1980cites this paper
• Estrogen synthetase. Effect of sulfhydryl reactive reagents on androgen aromatization by human term placental microsomes.

  1980cites this paper
• Induction of xenobiotic monooxygenases

  1980cites this paper
• Genetically mediated responses of microsomal ethanol oxidation in mice.

  1980cites this paper
• The history, properties, and function of NADPH-cytochrome P-450 reductase.

  1980cites this paper
• Ultrastructural and biochemical changes induced by progressive lipid peroxidation on isolated microsomes and rat liver endoplasmic reticulum.

  1980cites this paper
• Phospholipid asymmetry in rough- and smooth-endoplasmic-reticulum membranes of untreated and phenobarbital-treated rat liver.

  1980cites this paper
• Gram-Negative Endotoxin Administration Decreases Hepatic Drug-Metabolizing Enzymes during Development in Rats

  1980cites this paper
• Altered regulation of hepatic heme metabolism by indium chloride.

  1979cites this paper
• The role of cytochrome P-450 forms in 2-aminoanthracene and benz[α]pyrene mutagenesis

  1979cites this paper
• N-demethylation of aminopyrine in vivo and in the isolated hepatocyte of the rat.

  1979cites this paper
• Interaction of a series of beta-adrenergic blocking drugs with rat hepatic microsomal monooxygenase.

  1979cites this paper
• The role of the stimulation of nadph-cytochrome P-450 reductase activity in hepatic, microsomal mixed function oxidase activity.

  1979cites this paper
• The role of cytochrome P-450 forms in 2-aminoanthracene and benz[alpha]pyrene mutagenesis.

  1979cites this paper
• Microsomal drug hydroxylase activity of Tetrahymena pyriformis.

  1979cites this paper
• Glucose dehydrogenase (hexose 6-phosphate dehydrogenase) and the microsomal electron transport system. Evidence supporting their possible functional relationship.

  1979cites this paper
• Hepatic drug metabolism in pseudohermaphroditic rats.

  1979cites this paper
• In vitro metabolism of retinoic acid in hamster intestine and liver.

  1979cites this paper
• Stoichiometry of the mixed function oxidase.

  1979cites this paper
• Early biochemical liver changes following thiobenzamide poisoning.

  1979cites this paper
• Biological activity of methylglyoxal and related aldehydes.

  1978cites this paper
• Altered regulation of mammalian hepatic heme biosynthesis and urinary porphyrin excretion during prolonged exposure to sodium arsenate.

  1978cites this paper
• Ecdysterone biosynthesis: a microsomal cytochrome-P-450-linked ecdysone 20-monooxygenase from tissues of the African migratory locust.

  1978cites this paper
• Alterations in hepatic heme biosynthetic capability and mixed function oxidase activity during ethionine exposure in rats.

  1978cites this paper
• Agents activating the liver microsomal mixed function oxidase system

  1978cites this paper
• Effect of ethylenebisdiisothiocyanato sulfide (EBIS) on hepatic monooxygenase activity.

  1978cites this paper
• Pyridine nucleotide involvement in rat hepatic microsomal drug metabolism--I. Factors that influence NADPH kinetic estimations during mixed function oxidase reactions.

  1978cites this paper
• Spin-trapping studies of hydroxyl radical production involved in lipid peroxidation.

  1978cites this paper
• Hydroxylation of prostaglandins A1 and E1 by liver microsomal monooxygenase. Characteristics of the enzyme system in the guinea pig.

  1978cites this paper
• Effects of ischemia on drug-metabolizing microsomal enzymes in rat liver.

  1978cites this paper
• Carbon tetrachloride-induced lipid peroxidation of rat liver microsomes in vitro.

  1977cites this paper
• Spin trapping and its application in the study of lipid peroxidation and free radical production with liver microsomes.

  1977cites this paper
• Hepatic aminopyrine N-demethylase system: further studies of assay procedure.

  1977cites this paper
• CHARACTERISTICS OF THE HEPATIC MONOOXYGENASE CATALYZING THE METABOLISM OF PROSTAGLANDIN A1 (PGA1) IN GUINEA PIGS AND RATS. I. EFFECTS OF INDUCERS

  1977cites this paper
• The effects of phenobarbital and 3,4-benzypyrene on microsomal cytochrome P-450 and NADPH-cytochrome C reductase in regenerating rat liver after partial hepatectomy or chemical injury.

  1977cites this paper
• Studies on three microsomal electron transfer enzyme systems. Specificity of electron flow pathways.

  1977cites this paper
• PURIFICATION OF MEMBRANE–BOUND OXYGENASES: ISOLATION OF TWO ELECTROPHORETICALLY HOMOGENEOUS FORMS OF LIVER MICROSOMAL CYTOCHROME P-450

  1977cites this paper
• Effect of cholesterol feeding on tissue lipid perioxidation, glutathione peroxidase activity and liver microsomal functions in rats and guinea pigs.

  1977cites this paper
• Induction of liver lysosomal enzymes during the autophagic phase following phenobarbital treatment of rat

  1977cites this paper
• Studies on the N-demethylation and O-de-ethylation of ethylmorphine by hepatic microsomes from male rats.

  1977cites this paper
• Liver microsomal DT diaphorase: noninvolvement in hydroxylation of benzo(a)pyrene.

  1977cites this paper
• The effect of temperature on monoxygenase reactions in the microsomal membrane.

  1977cites this paper
• Role of sulfhydryl gropus in the hydroxylation of benzo[a]pyrene by purified rat and rabbit cytochrome P-448.

  1977cites this paper