Alcohol dehydrogenase-coupled spectrophotometric assay of epoxide hydratase activity

Guengerich, F. Peter; Mason, P S

doi:10.1016/0003-2697(80)90097-4

Cited by 27 publications

(6 citation statements)

References 17 publications

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“…Epoxide Hydrolase (EH) Activities. EH activities in vascular SMCs were determined spectrophotometrically by a coupled assay (Guengerich and Mason, 1980;Moorthy and Randerath, 1997) that measured the ability of aldehyde dehydrogenase to transfer electrons from styrene-7,8-dihdrodiol to NAD ϩ , formed as a result of EHcatalyzed hydrolysis of styrene-7,8 oxide. The EH activities were measured in the same cell pellets that were used for determining CYP1B1 expression.…”

Section: Methodsmentioning

confidence: 99%

Role of Cytochrome P4501B1 in Benzo[a]pyrene Bioactivation to DNA-Binding Metabolites in Mouse Vascular Smooth Muscle Cells: Evidence from³²P-Postlabeling for Formation of 3-Hydroxybenzo[a]pyrene and Benzo[a]pyrene-3,6-quinone as Major Proximate Genotoxic Intermediates

Moorthy¹,

Miller²,

Jiang³

et al. 2003

J Pharmacol Exp Ther

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Benzo [a]pyrene (BP), a polycylic aromatic hydrocarbon (PAH), is a potent atherogen and carcinogen in laboratory animals. Since genotoxic mechanisms may contribute to the development of atherosclerosis by PAHs, we have tested the hypotheses that: 1) BP induces DNA adducts in mouse aortic smooth muscle cells (SMCs); 2) 3-hydroxybenzo[a]pyrene (3-OH-BP) and benzo [a]pyrene-3,6-quinone (BPQ) are proximate genotoxic metabolites; and 3) cytochrome P4501B1 (CYP1B1) mediates the activation of BP and its metabolites to ultimate genotoxic intermediates. Cultured mouse aortic SMCs were treated with BP, 3-OH-BP, or BPQ for 24 h, and DNA adduct formation was analyzed by 32 P-postlabeling. In some experiments, cells were pretreated with the CYP1B1 inhibitor 1-ethynylpyrene (EP) prior to exposure to BP or its metabolites. BP, 3-OH-BP, and BPQ induced formation of several DNA adducts that were not observed in dimethylsulfoxide-treated cells. Reand cochromatography experiments indicated that 3-OH-BP and BPQ were proximate genotoxic metabolites of BP. DNA adduct formation was strongly inhibited by EP, a specific inhibitor of CYP1B1. BP treatment of SMCs resulted in induction of aryl hydrocarbon hydroxylase (AHH) activity and CYP1B1, but not CYP1A1, apoprotein. EP also blocked AHH induction by BP. In conclusion, the results of this study support the hypothesis that in SMCs, which are target sites for the development of atherosclerosis, the major bioactivation pathway of BP entails CYP1B1-mediated formation of the 3-OH-BP and BPQ, which are proximate genotoxic metabolites that may in turn get transformed to ultimate DNA-binding metabolites, which may contribute to atherogenesis by PAHs.

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Section: Methodsmentioning

confidence: 99%

Role of Cytochrome P4501B1 in Benzo[a]pyrene Bioactivation to DNA-Binding Metabolites in Mouse Vascular Smooth Muscle Cells: Evidence from³²P-Postlabeling for Formation of 3-Hydroxybenzo[a]pyrene and Benzo[a]pyrene-3,6-quinone as Major Proximate Genotoxic Intermediates

Moorthy¹,

Miller²,

Jiang³

et al. 2003

J Pharmacol Exp Ther

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“…The cytochrome P450-dependent oxidation of xenobiotics in hepatic microsomes uses an electron transport pathway to deliver reducing equivalents from Guengerich and Mason (1980) Note. Additional information on in vitro, phase I drug metabolism assays (and purification of the enzymes) can be found in Guengerich, F.P.…”

Section: 1 Experiments 1 Cofactor Requirements Of Drug Metabolismmentioning

confidence: 99%

Introduction to Drug Metabolism

Gibson

Skett

1994

201

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A catalogue record for this book is available from the British Library Library of Congress Catalog Card Number: 93-73495Printed on acid-free text paper, manufactured in accordance with ANSI/NISO Z39.48-1992 and ANSI/NISO Z39.48-1984 (Permanence of Paper). v Preface to second editionIt is now eight years since the publication of the first edition of Introduction to Drug Metabolism and much has changed in the world of drug biotransformation.The routes of metabolism are still relevant but much new information is available about the enzymology of the reactions (including a completely new method of naming many of the enzyme isoforms), the control mechanisms and methods of studying drug metabolism. We felt, therefore, that it was time to update the text to incorporate these new ideas.The textbook is intended to be a primer in drug metabolism for advanced undergraduate classes in science, medicine, dentistry and pharmacy, and for postgraduate students new to the area of drug metabolism. It is also useful as a source book of methods for undergraduate experiments in drug metabolism.The layout and content of the text is little changed from the first edition, as this was well received and we do not believe in tampering with something that is working. We intended to show the routes of metabolism first (chapter 1) and then further describe the enzymology of the reactions (chapter 2), followed by the important phenomena of induction and inhibition (chapter 3). The lessons learned in the first three chapters are used to illustrate how factors can affect drug metabolism (chapters 4 and 5) and the pharmacological, toxicological and clinical significance of drug metabolism (chapters 6 and 7). Chapter 8 is the methods section for a menu of drug metabolism related practicals.The expertise of the two authors is complementary, with one based on biochemistry and toxicology and the other on pharmacology and medicine. This is reflected in the balanced approach to the subject in this text. There is no other textbook at this level on drug metabolism -students have recourse only to more advanced texts, monographs and review articles. Many of these are given as a further reading list at the end of each chapter.We are indebted to our many colleagues and former students for comments on the first edition which have helped to make the second edition, we believe,

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“…Various epoxides Spectrophotometric Guengerich and Mason (1980) Note Additional information on in vitro. phase I drug metabolism assays (and purification of the enzymes) can be found in .…”

Section: Epoxide Hydrationmentioning

confidence: 99%

Introduction to Drug Metabolism

Gibson¹,

Skett²

1986

198

215

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This title is available in both hardbound and paperback editions. The paperback edition is sold subject to the condition that it shall not, by way of trade or otherwise, be lent, resold, hired out, or otherwise circulated without the publisher's prior consent in any form of binding or cover other than that in which it is published and without a similar condition including this condition being imposed on the subsequent purchaser.All rights reserved. No part of this book may be reprinted, or reproduced or utilized in any form or by an electronic, mechanical or other means, now PrefaceAlthough the scientific literature on drug metabolism is extensive, it suffers from the disadvantage that the material is diffuse and consists largely of specialist monographs dealing with particular aspects of the subject. In addition, although there are a few excellent texts on drug metabolism in print, these tend to be earlier publications and hence do not take into account the many recent advances in this area. Our motivations for writing this book therefore arose from the clear need for a recent and cohesive introductory text on this subject, specifically designed to cater for the needs of undergraduate and postgraduate students. Much of the subject matter in this text is derived from various courses on drug metabolism given at the University of Surrey and the University of Glasgow to basic science students in pharmacology, biochemistry, nutrition and nursing studies, to pre-clinical medical students and to undergraduate and post-graduate students in toxicology. Therefore, it is our intention that this text will serve as a primer in drug metabolism to a variety of students in the life sciences taking courses in this subject.The term 'drug metabolism' in its broadest sense may be considered as the absorption, distribution, biotransformation and excretion of drugs. To cover all these facets of drug metabolism in a single text is a voluminous task and therefore we have focused primarily on the biotransformation aspects of the subject. Having said this, the text is not solely a list of drug metabolism pathways, but rather it uses biotransformation reactions to rationalize many ix some of their published material. Finally, we express our thanks to the staff of Chapman and Hall for their excellent co-operation and for giving us the opportunity to bring this project to fruition.

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Alcohol dehydrogenase-coupled spectrophotometric assay of epoxide hydratase activity

Cited by 27 publications

References 17 publications

Role of Cytochrome P4501B1 in Benzo[a]pyrene Bioactivation to DNA-Binding Metabolites in Mouse Vascular Smooth Muscle Cells: Evidence from³²P-Postlabeling for Formation of 3-Hydroxybenzo[a]pyrene and Benzo[a]pyrene-3,6-quinone as Major Proximate Genotoxic Intermediates

Role of Cytochrome P4501B1 in Benzo[a]pyrene Bioactivation to DNA-Binding Metabolites in Mouse Vascular Smooth Muscle Cells: Evidence from³²P-Postlabeling for Formation of 3-Hydroxybenzo[a]pyrene and Benzo[a]pyrene-3,6-quinone as Major Proximate Genotoxic Intermediates

Introduction to Drug Metabolism

Introduction to Drug Metabolism

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Alcohol dehydrogenase-coupled spectrophotometric assay of epoxide hydratase activity

Cited by 27 publications

References 17 publications

Role of Cytochrome P4501B1 in Benzo[a]pyrene Bioactivation to DNA-Binding Metabolites in Mouse Vascular Smooth Muscle Cells: Evidence from32P-Postlabeling for Formation of 3-Hydroxybenzo[a]pyrene and Benzo[a]pyrene-3,6-quinone as Major Proximate Genotoxic Intermediates

Role of Cytochrome P4501B1 in Benzo[a]pyrene Bioactivation to DNA-Binding Metabolites in Mouse Vascular Smooth Muscle Cells: Evidence from32P-Postlabeling for Formation of 3-Hydroxybenzo[a]pyrene and Benzo[a]pyrene-3,6-quinone as Major Proximate Genotoxic Intermediates

Introduction to Drug Metabolism

Introduction to Drug Metabolism

Contact Info

Product

Resources

About

Role of Cytochrome P4501B1 in Benzo[a]pyrene Bioactivation to DNA-Binding Metabolites in Mouse Vascular Smooth Muscle Cells: Evidence from³²P-Postlabeling for Formation of 3-Hydroxybenzo[a]pyrene and Benzo[a]pyrene-3,6-quinone as Major Proximate Genotoxic Intermediates

Role of Cytochrome P4501B1 in Benzo[a]pyrene Bioactivation to DNA-Binding Metabolites in Mouse Vascular Smooth Muscle Cells: Evidence from³²P-Postlabeling for Formation of 3-Hydroxybenzo[a]pyrene and Benzo[a]pyrene-3,6-quinone as Major Proximate Genotoxic Intermediates