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NADPH-cytochrome P-450 reductase from rat liver: purification by affinity chromatography and characterization
Abstract
(NADPH)-cytochrome P-450 reductase was purified to apparent homogeneity by a procedure utilizing nicotinamide adenine dinucleotide phosphate (NADP)-Sepharose affinity column chromatography. The purified flavoprotein has a molecular weight of 79 700 and catalyzes cytochrome P-450 dependent drug metabolism, as well as reduction of exogenous electron acceptors. Aerobic titration of cytochrome P-450 reductase with NADPH indicates that an air-stable reduced form of the enzyme is generated by the addition of 0.5 mol of NADPH per mole of flavin, as judged by spectral characteristics. Further addition of NADPH causes no other changes in the absorbance spectrum. A Km value for NADPH of 5 micron was observed when either cytochrome P-450 or cytochrome c was employed as electron acceptor. A Km value of 8 +/- 2 micron was determined for cytochrome c and a Km of 0.09 +/- 0.01 micron was estimated for cytochrome P-450.
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Cited by
4-Hydroxylation of estrogens as marker of human mammary tumors.
Liehr, J. G., & Ricci, M. J. (1996) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.93.8.3294 · Cited by 414
[7] Purification and properties of NADPH-Cytochrome P-450 reductase
Strobel, H. W., & Dignam, J. D. (1978) · Methods in Enzymology · doi:10.1016/s0076-6879(78)52009-0 · Cited by 297
Self-citationIlan, Z., Ilan, R., & Cinti, D. L. (1981) · Journal of Biological Chemistry · doi:10.1016/s0021-9258(19)68742-9 · Cited by 75
Metadata
- Authors
- Dignam, J. D., & Strobel, H. W.
- Venue
- Biochemistry
- Publisher
- American Chemical Society (ACS)
- Published
- 1977-03-22
- Volume
- 16
- Issue
- 6
- Pages
- 1116-1123
- ISSN
- 0006-2960 (ISSN-L)
- 1520-4995
- OpenAlex
- W2072291087
- Cited by
- 206
- References
- 0
- FWCI
- 13.91
- Type
- journal article
- Language
- en
- Open access
- closed
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- Dignam, J. D., & Strobel, H. W. (1977). NADPH-cytochrome P-450 reductase from rat liver: purification by affinity chromatography and characterization. Biochemistry. https://doi.org/10.1021/bi00625a014
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