Cytochrome P450 Protocols / Edition 2

Cytochrome P450 Protocols / Edition 2

by Ian R. Phillips
     
 

Cytochromes P450 (CYPs) comprise a large superfamily of proteins that are of central importance in the detoxification or activation of a tremendous number of natural and synthetic hydrophobic xenobiotics, including many therapeutic drugs, chemical carcinogens and environmental pollutants. CYPs are important in mediating interactions between an organism and its

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Overview

Cytochromes P450 (CYPs) comprise a large superfamily of proteins that are of central importance in the detoxification or activation of a tremendous number of natural and synthetic hydrophobic xenobiotics, including many therapeutic drugs, chemical carcinogens and environmental pollutants. CYPs are important in mediating interactions between an organism and its chemical environment and in the regulation of physiological processes. Cytochrome P450 Protocols, Third Edition focuses on high-throughput methods for the simultaneous analysis of multiple CYPs, substrates or ligands. Although the emphasis is on CYPs of mammalian origin, it reflects an increasing interest in CYPs of bacterial species. Also included are chapters on cytochrome P450 reductase (the redox partner of CYPs) and the flavin-containing monooxygenases (FMOs), and metabolomic and lipidomic approaches for identification of endogenous substrates of CYPs (‘de-orphanizing’ CYP substrates). Written in the successful Methods in Molecular Biology™ series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls.

Authoritative and easily accessible, Cytochrome P450 Protocols, Third Edition provides a wide range of techniques accessible to researchers in fields as diverse as biochemistry, molecular biology, pharmacology, toxicology, environmental biology and genetics.

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Product Details

ISBN-13:
9781588294418
Publisher:
Springer-Verlag New York, LLC
Publication date:
12/15/2005
Series:
Methods in Molecular Biology Series, #320
Edition description:
2nd ed. 2006
Pages:
364
Product dimensions:
9.21(w) x 6.14(h) x 0.88(d)

Table of Contents

1. Bioluminescent Assays for Cytochrome P450 Enzymes

Douglas S. Auld, Henrike Veith, and James J. Cali

2. In Vitro Drug-Drug Interaction Assay

Michael Zientek and Kuresh Youdim

3. High-Throughput Mass Spectrometric Cytochrome P450 Inhibition Screening

Kheng B. Lim, Can C. Ozbal, and Daniel B. Kassel

4. The Synthesis, Characterization and Application of 13C-Methyl Isocyanide as an NMR Probe of Heme Protein Active Sites

Christopher McCullough, Phani Kumar Pullela, Sang-Choul Im, Lucy Waskell, Daniel Sem

5. High-Throughput Fluorescence Assay for Cytochrome P450 Mechanism-Based Inactivators

Cesar Kenaan, Haoming Zhang, and Paul F. Hollenberg

6. Identification of Endogenous Substrates of Orphan Cytochrome P450 Enzymes Through the Use of Untargeted Metabolomics Approaches

Qian Cheng and F. Peter. Guengerich

7. Genetic and Mass Spectrometric Tools for Elucidating the Physiological Function(s) of Cytochrome P450 Enzymes from Mycobacterium Tuberculosis

Hugues Ouellet, Eric D. Chow, Shenheng Guan, Jeffery S. Cox, Alma L. Burlingame, and Paul R. Ortiz de Montellano

8. An Escherichia coli Expression-Based Approach for Porphyrin Substitution in Heme Proteins

Michael B. Winter, Joshua J. Woodward, and Michael A. Marletta

9. Expression in Escherichia coli of a Cytochrome P450 Enzyme with a Cobalt Protoporphyrin IX Prosthetic Group

Wesley E. Straub, Clinton R. Nishida, and Paul R. Ortiz de Montellano

10. Nanodiscs in the Studies of Membrane-Bound Cytochrome P450 Enzymes

A. Luthra, M. Gregory, Y.V. Grinkova, I.G. Denisov , and S.G. Sligar

11. Rapid LC-MS Drug Metabolite Profiling Using Bioreactor Particles

Linlin Zhao, Besnik Bajrami, and James F. Rusling

12. Fluorescence-Based Screening of cytochrome P450 activities in Intact Cells

M. Teresa Donato and M. José Gómez-Lechón

13. Screening for Cytochrome P450 Reactivity with a Reporter Enzyme

Kersten S. Rabe and Christof M. Niemeyer

14. High-Throughput Fluorescence Assay of Cytochrome P450 3A4

Qian Cheng and F. Peter. Guengerich

15. Targeted Protein Capture for Analysis of Electrophile-Protein Adducts

Rebecca E. Connor, Simona G. Codreanu, Lawrence J. Marnett, and Daniel C. Liebler

16. DNA Shuffling of Cytochrome P450 Enzymes

James B.Y.H. Behrendorff, Wayne A. Johnston, and Elizabeth M.J. Gillam

17. Measurement of P450 Difference Spectra Using Intact Cells

Wayne A. Johnston and Elizabeth M.J. Gillam

18. DNA Shuffling of Cytochromes P450 for Indigoid Pigment Production

Nedeljka N. Rosic

19. P450 oxidoreductase – Genotyping, Expression, Purification of Recombinant Protein and Activity Assessments of Wild-Type and Mutant Protein

Vishal Agrawal and Walter L. Miller

20. LICRED: A Versatile Drop-In Vector for Rapid Generation of Redox-Self-Sufficient Cytochromes P450

Federico Sabbadin, Gideon G. Grogan and Neil C. Bruce

21. Update on Allele Nomenclature for Human Cytochromes P450 and the Human Cytochrome P450 Allele (CYP-allele) Nomenclature Database

Sarah C. Sim and Magnus Ingelman-Sundberg

22. Simultaneous in vivo Phenotyping of CYP Enzymes

Sussan Ghassabian and Michael Murray

23. Detection of Regulatory Polymorphisms: High-Throughput Capillary DNase-I Footprinting

Matthew Hancock and Elizabeth A. Shephard

24. Isolation of Mouse Hepatocytes

Mina Edwards, Lyndsey Houseman, Ian R. Phillips, and Elizabeth A. Shephard

25. Highly Efficient SiRNA and Gene Transfer into Hepatocyte-like HepaRG Cells and Primary Human Hepatocytes: New Means for Drug Metabolism and Toxicity Studies

Véronique Laurent, Denise Glaise, Tobias Nübel, David Gilot, Anne Corlu, and Pascal Loyer

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