Radiometric assay for cytochrome P-450-catalyzed progesterone 16[alpha]-hydroxylation and determination of an apparent isotope effect
dc.contributor.author | Osawa, Yoichi | en_US |
dc.contributor.author | Coon, Minor J. | en_US |
dc.date.accessioned | 2006-04-07T19:50:23Z | |
dc.date.available | 2006-04-07T19:50:23Z | |
dc.date.issued | 1987-08-01 | en_US |
dc.identifier.citation | Osawa, Yoichi, Coon, Minor J. (1987/08/01)."Radiometric assay for cytochrome P-450-catalyzed progesterone 16[alpha]-hydroxylation and determination of an apparent isotope effect." Analytical Biochemistry 164(2): 355-361. <http://hdl.handle.net/2027.42/26627> | en_US |
dc.identifier.uri | http://www.sciencedirect.com/science/article/B6W9V-4DVNP0N-T7/2/ea69286d4b694ced1dec739e6a534619 | en_US |
dc.identifier.uri | https://hdl.handle.net/2027.42/26627 | |
dc.identifier.uri | http://www.ncbi.nlm.nih.gov/sites/entrez?cmd=retrieve&db=pubmed&list_uids=3674383&dopt=citation | en_US |
dc.description.abstract | In the course of studies on the oxygenation of steroids by purified P-450 cytochromes, particularly rabbit liver microsomal cytochrome P-450 form 3b, a rapid and reliable radiometric assay has been devised for progesterone 16[alpha]-hydroxylation. In view of the lack of a commercially available, suitably tritiated substrate, [1,2,6,7,16,17-3H]progesterone was treated with alkali to remove the label from potential hydroxylation sites other than the 16[alpha] position. The resulting [1,7,16-3H]progesterone was added to a reconstituted enzyme system containing cytochrome P-450 form 3b, NADPH-cytochrome P-450 reductase, and NADPH, and the rate of 16[alpha]-hydroxylation was measured by the formation of 3H2O. This reaction was shown to be linear with respect to time and to the cytochrome P-450 concentration. An apparent tritium isotope effect of 2.1 was observed by comparison of the rates of formation of tritium oxide and 16[alpha]-hydroxyprogesterone, and the magnitude of the isotope effect was confirmed by an isotope competition assay in which a mixture of [1,7,16-3H]progesterone and [4-14C]progesterone was employed. | en_US |
dc.format.extent | 594987 bytes | |
dc.format.extent | 3118 bytes | |
dc.format.mimetype | application/pdf | |
dc.format.mimetype | text/plain | |
dc.language.iso | en_US | |
dc.publisher | Elsevier | en_US |
dc.title | Radiometric assay for cytochrome P-450-catalyzed progesterone 16[alpha]-hydroxylation and determination of an apparent isotope effect | en_US |
dc.type | Article | en_US |
dc.rights.robots | IndexNoFollow | en_US |
dc.subject.hlbsecondlevel | Public Health | en_US |
dc.subject.hlbsecondlevel | Chemistry | en_US |
dc.subject.hlbsecondlevel | Chemical Engineering | en_US |
dc.subject.hlbsecondlevel | Biological Chemistry | en_US |
dc.subject.hlbtoplevel | Engineering | en_US |
dc.subject.hlbtoplevel | Science | en_US |
dc.subject.hlbtoplevel | Health Sciences | en_US |
dc.description.peerreviewed | Peer Reviewed | en_US |
dc.contributor.affiliationum | Department of Pharmacology, Medical School, The University of Michigan, Ann Arbor, Michigan 48109, USA; Department of Biological Chemistry, Medical School, The University of Michigan, Ann Arbor, Michigan 48109, USA. | en_US |
dc.contributor.affiliationum | Department of Pharmacology, Medical School, The University of Michigan, Ann Arbor, Michigan 48109, USA; Department of Biological Chemistry, Medical School, The University of Michigan, Ann Arbor, Michigan 48109, USA. | en_US |
dc.identifier.pmid | 3674383 | en_US |
dc.description.bitstreamurl | http://deepblue.lib.umich.edu/bitstream/2027.42/26627/1/0000168.pdf | en_US |
dc.identifier.doi | http://dx.doi.org/10.1016/0003-2697(87)90504-5 | en_US |
dc.identifier.source | Analytical Biochemistry | en_US |
dc.owningcollname | Interdisciplinary and Peer-Reviewed |
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