Preface ......................................................... v
Contributors ................................................... ix
1 Bioluminescent Assays for Cytochrome P450 Enzymes ............ 1
Douglas S. Auld, Henrike Veith, and James J. Cali
2 Simultaneous Determination of Multiple CYP Inhibition
Constants using a Cocktail-Probe Approach ................... 11
Michael Zientek and Kuresh Youdim
3 High-Throughput Mass Spectrometric Cytochrome P450
Inhibition Screening ........................................ 25
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 ................................................ 51
Christopher McCullough, Phani Kumar Pullela, Sang-Choul Im,
Lucy Waskell, and Daniel Sem
5 High-Throughput Fluorescence Assay for Cytochrome P450
Mechanism Based Inactivators ................................ 61
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 ..................................... 71
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 ............................. 79
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 ..................... 95
Michael B. Winter, Joshua J. Woodward, and Michael
A. Marietta
9 Expression in Escherichia coli of a Cytochrome P450 Enzyme
with a Cobalt Protoporphyrin IX Prosthetic Group ........... 107
Wesley E. Straub, Clinton R. Nishida, and Paul R Ortiz de
Montellano
10 Nanodiscs in the Studies of Membrane-Bound Cytochrome
P450 Enzymes ............................................... 115
A. Luthra, M. Gregory, Y.V. Grinkova, I.G. Denisov, and
S.G. Sligar
11 Rapid LC-MS Drug Metabolite Profiling Using Bioreactor
Particles .................................................. 129
Linlin Zhao, Besnik Bajrami, and James F. Rusting
12 Fluorescence-Based Screening of Cytochrome P450 Activities
in Intact Cells ............................................ 135
M. Teresa Donate and M. José Gómez-Lechón
13 Screening for Cytochrome P450 Reactivity with a Reporter
Enzyme ..................................................... 149
Kersten S. Rabe and Christof M. Niemeyer
14 High Throughput Fluorescence Assay of Cytochrome P450 3A4 .. 157
Qian Cheng and F. Peter Guengerich
15 Targeted Protein Capture for Analysis of Electrophile-
Protein Adducts ............................................ 163
Rebecca E. Connor, Simona G. Codreanu, Lawrence J.
Marnett, and Daniel C. Liebler
16 DNA Shuffling of Cytochrome P450 Enzymes ................... 177
James B.T.H. Bebrendorff, Wayne A. Johnston, and
Elizabeth M.J. Gillam
17 Measurement of P450 Difference Spectra Using Intact Cells .. 189
Wayne A. Johnston and Elizabeth M.J. Gillam
18 DNA Shuffling of Cytochromes P450 for Indigoid Pigment
Production ................................................. 205
Nedeljka N. Rosic
19 P450 Oxidoreductase: Genotyping, Expression, Purification
of Recombinant Protein, and Activity Assessments of Wild-
Type and Mutant Protein .................................... 225
Vishal Agrawal and Walter L. Miller
20 LICRED: A Versatile Drop-In Vector for Rapid Generation
of Redox-Self-Sufficient Cytochromes P450 .................. 239
Federico Sabbadin, Gideon Grogan, and Neil C. Bruce
21 Update on Allele Nomenclature for Human Cytochromes P450
and the Human Cytochrome P450 Allele (CYP-Allele)
Nomenclature Database ...................................... 251
Sarah C. Sim and Magnus Ingelman-Sundberg
22 Simultaneous In Vivo Phenotyping of CYP Enzymes ............ 261
Sussan Ghassabian and Michael Murray
23 Detection of Regulatory Polymorphisms: High-Throughput
Capillary DNase I Footprinting ............................. 269
Matthew Hancock and Elizabeth A. Shephard
24 Isolation of Mouse Hepatocytes ............................. 283
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 ................... 295
Véronique Laurent, Denise Glaise, Tobias Nülbel, David
Gilot, Anne Corlu, and Pascal Loyer
Index ......................................................... 315
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