Measurement of cytochrome P450 and NADPH-cytochrome P450 reductase

被引：313

作者：

Guengerich, F. Peter ^{[1
]}

Martin, Martha V.

Sohl, Christal D.

Cheng, Qian

机构：

[1] Vanderbilt Univ, Sch Med, Dept Biochem, Nashville, TN 37212 USA

来源：

NATURE PROTOCOLS | 2009年 / 4卷 / 09期

关键词：

ALLENE OXIDE SYNTHASE; RAT-LIVER MICROSOMES; P-450; REDUCTASE; SPECTRAL CHARACTERIZATION; DRUG-INTERACTIONS; ESCHERICHIA-COLI; HUMAN AROMATASE; C REDUCTASE; PURIFICATION; OXIDATION;

D O I：

10.1038/nprot.2009.121

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Cytochrome P450 (P450) enzymes are important in the metabolism of steroids, vitamins, carcinogens, drugs and other compounds. Two of the commonly used assays in this field are the measurements of total P450 and NADPH-P450 reductase in biological preparations. A detailed protocol is presented for the measurement of P450 by its spectral properties, along with a protocol for measuring NADPH-P450 reductase by its NADPH-cytochrome c reduction activity. Each assay can be completed in 5-10 min. Detailed explanations for the rationale of particular sequences in the protocols are provided, along with potential confounding problems.

引用

页码：1245 / 1251

页数：7

共 32 条

[1] High-throughput fluorescence assay of cytochrome P450 3A4 [J].

Cheng, Qian ;

Sohl, Christal D. ;

Guengerich, F. Peter .

NATURE PROTOCOLS, 2009, 4 (09) :1258-1261

[2]

Coon M.J., 1976, STRUCTURAL BASIS MEM, P409, DOI DOI 10.1016/B978-0-12-332450-4.50035-4

[3] PREPARATION OF HOMOGENEOUS NADPH-CYTOCHROME P-450 REDUCTASE FROM RAT-LIVER [J].

DIGNAM, JD ;

STROBEL, HW .

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1975, 63 (04) :845-852

[4] Human aromatase in high yield and purity by perfusion chromatography and its characterization by difference spectroscopy and mass spectrometry [J].

Gartner, CA ;

Thompson, SJ ;

Rettie, AE ;

Nelson, SD .

PROTEIN EXPRESSION AND PURIFICATION, 2001, 22 (03) :443-454

[5] Structural basis for androgen specificity and oestrogen synthesis in human aromatase [J].

Ghosh, Debashis ;

Griswold, Jennifer ;

Erman, Mary ;

Pangborn, Walter .

NATURE, 2009, 457 (7226) :219-U119

[6] Extending the capabilities of nature's most versatile catalysts: Directed evolution of mammalian xenobiotic-metabolizing P450s [J].

Gillam, Elizabeth M. J. .

ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 2007, 464 (02) :176-186

[7] PURIFICATION OF CYTOCHROME-P-450, NADPH-CYTOCHROME P-450 REDUCTASE, AND EPOXIDE HYDRATASE FROM A SINGLE PREPARATION OF RAT-LIVER MICROSOMES [J].

GUENGERICH, FP ;

MARTIN, MV .

ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1980, 205 (02) :365-379

[8] Reduction of cytochrome b5 by NADPH-cytochrome P450 reductase [J].

Guengerich, FP .

ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 2005, 440 (02) :204-211

[9] DESTRUCTION OF HEME AND HEMOPROTEINS MEDIATED BY LIVER MICROSOMAL REDUCED NICOTINAMIDE ADENINE-DINUCLEOTIDE PHOSPHATE-CYTOCHROME P-450 REDUCTASE [J].

GUENGERICH, FP .

BIOCHEMISTRY, 1978, 17 (17) :3633-3639

[10]

Guengerich FP., 2005, Cytochrome P450: structure, metabolism, and biochemistry, V3rd, P377

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