Cytochrome P450: Volume 206: Protein-Dna Interactions


This volume represents the first collection of methodological approaches to P450 investigation which includes the use of molecular biological techniques. It is designed for use by those who study the P450 super family. Methods are presented for the identification and characterization of unique forms of P450 mRNA and protein, for expression of P450s in heterologous systems, for the study of human P450s, and for the characterization of P450s in different tissues.


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This volume represents the first collection of methodological approaches to P450 investigation which includes the use of molecular biological techniques. It is designed for use by those who study the P450 super family. Methods are presented for the identification and characterization of unique forms of P450 mRNA and protein, for expression of P450s in heterologous systems, for the study of human P450s, and for the characterization of P450s in different tissues.

Audience: Biochemists, cell biologists, molecular biologists, toxicologists, pharmacologists, microbiologists, immunologists, analytical chemists, biophysicists, endocrinologists, and graduate students in these disciplines.

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Editorial Reviews

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Praise for the Series
"The Methods in Enzymology series represents the gold-standard."
"Incomparably useful."
"It is a true 'methods' series, including almost every detail from basic theory to sources of equipment and reagents, with timely documentation provided on each page."
"The series has been following the growing, changing and creation of new areas of science. It should be on the shelves of all libraries in the world as a whole collection."
"The appearance of another volume in that excellent series, Methods in Enzymology, is always a cause for appreciation for those who wish to successfully carry out a particular technique or prepare an enzyme or metabolic intermediate without the tiresome prospect of searching through unfamiliar literature and perhaps selecting an unproven method which is not easily reproduced."
"If we had some way to find the work most often consulted in the laboratory, it could well be the multi-volume series Methods in Enzymology...a great work."
"A series that has established itself as a definitive reference for biochemists."
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Product Details

Table of Contents

Site-Directed Mutagenesis, Protein Modification, and Structural Analysis:
D.W. Nebert and D.R. Nelson, P450 Gene Nomenclature Based on Evolution.
T.L. Poulos, Modeling of Mammalian P450s on Basis of P450cam X-Ray Structure.
S.G. Sligar, D. Filipovic, and P.S. Stayton, Mutagenesis of Cytochromes P450cam and b5.
F. Millett and L.M. Geren, Chemical Modification of Interaction between Adrenodoxin and Cytochrome P450scc.
L.S. Kaminsky, D. Dunbar, and W. Lawson, Azidowarfarin as Photoaffinity Probe of Cytochromes P450.
E. Szczesna-Skorupa and B. Kemper, Cell-Free Analysis of Targeting of Cytochrome P450 to Microsomal Membranes.
T. Omura and A. Oto, Biosynthesis and Intracellular Sorting of Mitochondrial Forms of Cytochrome P450.
F.J. Gonzalez, T. Aoyama, and H.V. Gelboin, Expression of Mammalian Cytochrome P450 Using Vaccinia Virus.
F.J. Gonzalez, S. Kimura, S. Tamura, and H.V. Gelboin, Expression of Mammalian Cytochrome P450 Using Baculovirus.
B.J. Clark and M.R. Waterman, Heterologous Expression of Mammalian P450 cDNA in COS Cells.
T.D. Porter and J.R. Larson, Expression of Mammalian P450s in Escherichia coli.
J. Doehmer and F. Oesch, V79 Chinese Hamster Cells Genetically Engineered for Stable Expression of Cytochromes P450.
C.L. Crespi, Expression of Cytochrome P450 cDNAs in Human B Lymphoblastoid Cells: Applications to Toxicology and Metabolite Analysis.
F.P. Guengerich, W.R. Brian, M.-A. Sari, and J.T. Ross, Expression of Mammalian Cytochrome P450 Enzymes Using Yeast-Based Vectors.
Characterization of Genetic Polymorphism and Errors of Metabolism:
N.K. Spurr and C.R. Wolf, Genetic Analysis of Cytochrome P450 System.
Y. Higashi and Y. Fujii-Kuriyama, Functional Analysis of Mutant P450(C21) Genes in a COS Cell Expression System.
M.H. Heim and U.A. Meyer, Genetic Polymorphism of Debrisoquine Oxidation: Restriction Fragment Analysis and Allele-Specific Amplification of Mutant Alleles of CYP2D6.
R. Kato, T. Yasumori, and Y. Yamazoe, Characterization of Human P450IIC Isozyme Using Yeast Expression System.
Immunochemical Methods:
T. Friedberg, W. Kissel, M. Arand, and F. Oesch, Production of Site-Specific P450 Antibodies Using Recombinant Fusion Proteins as Antigens.
U.M. Zanger, Immunoisolation of Human Microsomal Cytochromes P450's Using Autoantibodies.
M.P. Manns and E.F. Johnson, Identification of Human Cytochrome P450 as Autoantigens.
R.J. Edwards, B.P. Murray, and A.R. Boobis, Antipeptide Antibodies in Studies of Cytochromes P450IA.
A. Parkinson and B. Gemzik, Production and Purification of Antibodies against Rat Liver P450 Enzymes.
Selective Measurement of Protein and mRNA Expression:
D.J. Waxman, Rat Hepatic P450IIA and P450IIC Subfamily Expression Using Catalytic, Immunochemical, and Molecular Probes.
M. Negishi, B. Burkhart, and K. Aida, Expression of Genes within Mouse IIA and IID Subfamilies: Simultaneous Measurement of Homologous P450 mRNAs.
J.P. Hardwick, CYP4A Subfamily: Functional Analysis by Immunohistochemistry and in Situ Hybridization.
R.H.~Tukey and S. Okino, Quantitation of Related Gene Products by Nuclear Run-On and Northern Blot Analysis.
C. Hassett, R. Ramsden, and C.J. Omiecinski, Isolating Cytochrome P450 cDNA and Genomic Clones: Library Screening with Synthetic DNA Oligomers.
Regulatory Paradigms:
J.A. Koch and D.J. Waxman, P450 Phosphorylation in Isolated Hepatocytes and in Vivo.
M.A. Correia, Cytochrome P450 Turnover.
J.B. Schenkman, Induction of Diabetes and Evaluation of Diabetic State on P450 Expression.
Cell Culture Systems:
D. Li, E.G. Schuetz, and P.S. Guzelian, Hepatocyte Culture in Study of P450 Regulation.
M. Daujat, L. Pichard, I. Fabre, T. Pineau, G. Fabre, C. Bonfils, and P. Maurel, Induction Protocols Cytochromes P450IIIA for in Vivo and in Primary Cultures of Animal and Human Hepatocytes.
G.G. Gibson and B.G. Lake, Induction Protocols for Cytochrome P450IVA Subfamily in Animals and Primary Hepatocyte Cultures.
D.R. Koop, R.M. Laethem, A.L. Goldner, and J.G. Douglas, Cytochrome P450 Expression and Metabolism Isolated Rabbit Renal Epithelium.
P.J. Hornsby and J.M. McAllister, Culturing Steroidogenic Cells.
O. Hankinson, Selection for and against Cells Possessing Cytochrome P450IA1-Dependent Aryl Hydrocarbon Hydroxylase Activity.
Transcriptional Regulation:
E.S. Shen, C.J. Elferink, and J.P. Whitlock, Jr. , Use of Gel Retardation to Analyze Protein-DNA Interactions Upstream of CYPIA1 Gene.
T.H. Rushmore and C.B. Pickett, Xenobiotic Responsive Elements Controlling Inducible Expression by Planar Aromatic Compounds and Phenolic Antioxidants.
Enzyme Assays:
A.D. Rodrigues and R.A. Prough, Induction of Cytochromes P450IA1 and P450IA2 and Measurement of Catalytic Activities.
R.T. Okita, J.E. Clark, J.R. Okita, and B.S.S. Masters, γu and (γu-1)-Hydroxylation of Eicosanoids and Fatty Acids by High-Performance Liquid Chromatography.
J.H. Capdevila, E. Dishman, A. Karara, and J.R. Falck, Cytochrome P450 Arachidonic Acid Epoxygenase: Stereochemical Characterization of Epoxyeicosatrienoic Acids.
M.P. Arlotto, J.M. Trent, and R.W. Estabrook, Measurement of Steroid Hydroxylation Reactions by HPLC as an Indicator of P450 Idenity and Function.
D.J. Waxman, P450-Catalyzed Steroid Hydroxylation: Assay and Product Identification by Thin-Layer Chromatography.
~E.D. Lephart and E.R. Simpson, Assay of Aromatase Activity.
J.Y.L. Chiang, Reversed-Phase HPLC Assay of Cholesterol 7αa-Hydroxylase.
J.L. Napoli, C.A. Martin, and R.L. Horst, Induction, Inhibition, and Analysis of Vitamin D Metabolism in Cultured Cells.
M.E. McManus and R.A. McKinnon, Measurement of Cytochrome P450 Activation of Xenobiotics Using Ames/Salmonella Test.
T. Kronbach, Bufuralol, Dextromethorphan, and Debrisoquine as Prototype Substrates for Human P450IID6.
P.B. Watkins, Breath Tests as Noninvasive Assays of P450s.
L.A. Waskell, J.L. Vigne, and G. Vergeres, Site of Action of Substrates Requiring Cytochrome b5 for Oxidation by Cytochrome P450.
Design and Characterization of Inhibitors:
P.R. Ortiz de Montellano, Mechanism-Based~Inactivation of Cytochrome P450: Isolation and Characterization of N-Alkyl Heme Adducts.
J.R. Halpert and J.C. Stevens, Dihalomethyl Compounds as Mechanism-Based Inactivators of Cytochromes P450.
L.E. Vickery, Active Site-Directed Inhibitors of Steroid Hydroxylase Cytochromes P450.
M.R. Franklin, Cytochrome P450 Metabolic or Intermediate Complexes from Macrolide Antibiotics and Related Compounds.
Isolation Procedures:
J.L. Raucy and J.M. Lasker, Isolation of P450 Enzymes from Human Liver.
R.M. Shayiq, S. Addya, and N.G. Avadhani, Constitutive and Inducible Forms of Cytochrome P450 from Hepatic Mitochondria.
C.S. Yang, C.J. Patten, H. Ishizaki, and J.-S.H. Yoo, Induction, Purification, and Characterization of Cytochrome P450IIE.
X. Ding and M.J. Coon, Purification and Characterization of Cytochromes P450 in Rabbit Nasal Microsomes.
J.A. Peterson and J.-Y. Lu, Bacterial Cytochromes P450: Isolation and Identification.
Characterization of Extrahepatic Forms of P450:
R.M. Philpot, Characterization of Cytochrome P450 in Extrahepatic Tissues.
M. Warner, R. Ahlgren, P.G. Zaphiropoulos, S.-I. Hayashi, and J.-A. Gustafsson, Identification and Localization of Cytochrones P450 Expressed in Brain.
M. Strömstedt, P.G. Zaphiropoulos, and J.-A. Gustafsson, Identification of Cytochrome P450 in Extrahepatic Tissues by Cross-Hybridization of Oligonucleotides and cDNAs.
H.W. Strobel, D.K. Hammond, T.B. White, and J.W. White, Identification and Localization of Cytochromes P450 in Gut.
Each chapter includes references.
Author Index.
Subject Index.

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