Cytochrome P450 Protocols / Edition 2

by Ian R. Phillips

Overview

For this second edition of their much praised Cyhrome P450, the editors have collected accounts of the essential core techniques that use the latest methodologies for the investigation of P450s. Highlights include prools for spectral analysis and purification of P450s, enzymatic assays of P450s and flavin-containing monooxygenases (FMOs), expression of P450s and FMOs in heterologous systems, and the production and use of antipeptide antibodies. Additional chapters contain readily reproducible techniques for the transfection of hepaytes for gene regulation studies, P450 reporter gene assays, in situ hybridization, and analysis of genetic polymorphisms. Although the emphasis is on P450s of mammalian origin, many of the readily reproducible methods described are suitable for P450s from any source.

Product Details

ISBN-13:	9781617375729
Publisher:	Springer-Verlag New York, LLC
Publication date:	11/19/2010
Series:	Methods in Molecular Biology , #320
Edition description:	2nd ed. 2006
Pages:	364
Product dimensions:	6.10(w) x 9.25(h) x 0.03(d)

About the Author

Phillips, Ian R. (Univ of London); Shepard, Elizabeth A. (Univ Coll London)

Table of Contents

Cyhrome P450 Nomenclature, 2004 David R. Nelson Spectral Analyses of Cyhromes P450 John B. Schenkman and Ingela Jansson Establishment of Functional Human Cyhrome P450 Monooxygenase Systems in Escherichia coli Michael P. Pritchard, Lesley McLaughlin, and Thomas Friedberg Purification of Cyhromes P450: Products of Bacterial Recombinant Expression Systems F. Peter Guengerich and Martha V. Martin Expression of Recombinant Flavin-Containing Monooxygenases in a Baculovirus/Insect Cell System Azara Janmohamed, Prapimpuk Thaunsukon, Elizabeth A. Shephard, and Ian R. Phillips Cyhrome P450 Reconstitution Systems Hiroshi Yamazaki and Tsutomu Shimada Catalytic Assays for Human Cyhrome P450: An Introduction Thomas K. H. Chang and David J. Waxman Enzymatic Analysis of cDNA-Expressed Human CYP1A1, CYP1A2, and CYP1B1 With 7-Ethoxyresorufin as Substrate Thomas K. H. Chang and David J. Waxman Spectrofluorometric Analysis of CYP2A6-Catalyzed Coumarin 7-Hydroxylation David J. Waxman and Thomas K. H. Chang Determination of CYP2B6 Component of 7-Ethoxy-4-Trifluoromethylcoumarin O-Deethylation Activity in Human Liver Microsomes Thomas K. H. Chang, Charles L. Crespi, and David J. Waxman High-Performance Liquid Chromatography Analysis of CYP2C8-Catalyzed Paclitaxel 6a-Hydroxylation Charles L. Crespi, Thomas K. H. Chang, and David J. Waxman Determination of CYP2C9-Catalyzed Diclofenac 4'-Hydroxylation by High-Performance Liquid Chromatography Charles L. Crespi, Thomas K. H. Chang, and David J. Waxman CYP2C19-Mediated (S)-Mephenytoin 4'-Hydroxylation Assayed by High-Performance Liquid Chromatography With Radiometric Detection Charles L. Crespi, Thomas K. H. Chang, and David J. Waxman CYP2D6-Dependent Bufuralol 1'-Hydroxylation Assayed by Reverse-Phase Ion-Pair High-Performance Liquid Chromatography With Fluorescence Detection Charles L. Crespi, Thomas K. H. Chang, and David J. Waxman Spectrophotometric Analysis of Human CYP2E1-Catalyzed p-Nitrophenol Hydroxylation Thomas K. H. Chang, Charles L. Crespi, and David J. Waxman Thin-Layer Chromatography Analysis of Human CYP3A-Catalyzed Testosterone 6b-Hydroxylation David J. Waxman and Thomas K. H. Chang Determination of CYP4A11-Catalyzed Lauric Acid 12-Hydroxylation by High-Performance Liquid Chromatography With Radiometric Detection Charles L. Crespi, Thomas K. H. Chang, and David J. Waxman An Isocratic High-Performance Liquid Chromatography Assay for CYP7A1-Catalyzed Cholesterol 7a-Hydroxylation David J. Waxman and Thomas K. H. Chang Use of 7-Ethoxycoumarin to Monitor Multiple Enzymes in the Human CYP1, CYP2, and CYP3 Families David J. Waxman and Thomas K. H. Chang Benzydamine N-Oxygenation as a Measure of Flavin-Containing Monooxygenase Activity Catherine K. Yeung and Allan E. Rettie Prochiral Sulfoxidation as a Probe for Flavin-Containing Monooxygenases Catherine K. Yeung and Allan E. Rettie Targeting Antipeptide Antibodies Toward Cyhrome P450 Enzymes Robert J. Edwards The Human Cyhrome P450 Allele Nomenclature Committee Web Site: Submission Criteria, Procedures, and Objectives Sarah C. Sim and Magnus Ingelman-Sundberg Genotyping for Cyhrome P450 Polymorphisms Ann K. Daly, Barry P. King, and Julian B. S. Leathart Hepayte Cultures in Drug Metabolism and Toxicological Research and Testing Tamara Vanhaecke and Vera Rogiers Isolation of Rat Hepaytes Peggy Papeleu, Tamara Vanhaecke, Tom Henkens, Greetje Elaut, Mathieu Vinken, Sarah Snykers, and Vera Rogiers Rat Hepayte Cultures: Conventional Monolayer and Cocultures With Rat Liver Epithelial Cells Tom Henkens,

Editorial Reviews

From Reviews of the First Edition...

"...a remarkable book, highly useful for all our scientific readers..." -Cellular and Molecular Biology

"...will be welcomed by laboratory scientists of various profiles who need authoritative prools for their studies." -Modern Pathology

From the reviews of the second edition:

"In this second edition, editors Phillips and Shephard have updated the text to reflect current advances and to include chapters on some more contemporary procedures … . I’m sure that the wealth of background information in the book would be valuable. Overall … this is a useful reference book for experienced researchers and for those new to this discipline. Recommended." (David Wilkinson, British Toxicology Society Newsletter, Winter, 2006)

From the Publisher

A collection of techniques for investigating the large superfamily of proteins that are of central importance in detoxifying or activating many foreign hydrophobic compounds, among them therapeutic drugs, chemical carcinogens, and environmental pollutants. The focus is on P450 species from mammals, but the methods are also suitable for those from other sources. The techniques described are used for purifying, spectral analysis, expression in heterologous cells, determining functional capabilities, detecting and quantifying, gene expression and its regulation, and cell culture. Addressed to biologists and chemists with no previous knowledge of the techniques. Annotation c. by Book News, Inc., Portland, Or.

Booknews

Reviewer: Jed N. Lampe, Ph.D.(University of Kansas Medical Center)
Description: With this third edition, the authors and contributors continue to provide what has become a tour de force of bench top manuals for the cytochrome P450 field. Building on the success of the first two editions, this book expands and diversifies a set of well-established, validated protocols for the production and analysis of cytochrome P450 enzymes (CYPs) and their diverse metabolic products.
Purpose: The authors focus on high-throughput techniques for the examination of multiple CYPs in both in vitro and in vivo models, which is a notable expansion and extension of the fundamental techniques established in the earlier two editions.
Audience: Although predominantly useful for an industrial audience due to the high-throughput nature of most of the techniques, this book should also prove of interest to graduate students and postdoctoral researchers who are new to the field of P450 science, or who would like to expand their research directions.
Features: The highlight of this edition, as well as the previous editions, is the step-by-step protocols with extensive troubleshooting notes at the end of each chapter. These are augmented with selected and appropriate color illustrations that enhance readers' understanding of each technique.
Assessment: This stands alone as the premier book for this field, which is sorely lacking in publications of this type. Researchers in the P450 field will find this to be a useful bench top guide that will provide them with the tools to explore and understand CYPs, both in vitro and in vivo.

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