| pubmed-article:2719667 | rdf:type | pubmed:Citation | lld:pubmed |
| pubmed-article:2719667 | lifeskim:mentions | umls-concept:C0039005 | lld:lifeskim |
| pubmed-article:2719667 | lifeskim:mentions | umls-concept:C0022646 | lld:lifeskim |
| pubmed-article:2719667 | lifeskim:mentions | umls-concept:C0006657 | lld:lifeskim |
| pubmed-article:2719667 | lifeskim:mentions | umls-concept:C0042890 | lld:lifeskim |
| pubmed-article:2719667 | pubmed:issue | 2 | lld:pubmed |
| pubmed-article:2719667 | pubmed:dateCreated | 1989-6-20 | lld:pubmed |
| pubmed-article:2719667 | pubmed:abstractText | Pig kidney mitochondrial 25-hydroxyvitamin D3 1 alpha- and 24-hydroxylase activities have been solubilized with cholate/Emulgen 911 and reconstituted with NADPH, ferredoxin reductase and ferredoxin. All three of these components are required for full catalytic activity of both enzymes. Both products were identified by co-chromatography with authentic metabolites on both normal and reverse-phase h.p.l.c. using solvent systems which were shown to separate 10-oxo-19-nor-25-hydroxyvitamin D3 from 1,25-dihydroxyvitamin D3 [1,25-(OH)2-D3]. In addition, periodate treatment of the 24,25-(OH)2-D3 product resulted in complete loss of the product as measured by protein-binding assay. Further purification by p-chloroamphetamine-Sepharose chromatography of a solubilized extract from a pig fed a normal diet increased the specific content of the cytochrome P-450 from 0.019 to 0.239 nmol/mg and the 1 alpha-hydroxylase activity from 4.75 to 268 pmol/h per mg. Activity of the 24-hydroxylase in the crude solubilized extract was 6.3 pmol/h per mg, but was undetectable after partial purification by a p-chloroamphetamine-Sepharose column. However, further fractionation of this material by DEAE-Sepharose chromatography resulted in a further increase in 1 alpha-hydroxylase activity to 430 pmol/h per mg and detection of 24-hydroxylase in a separate fraction at a level of 53 pmol/h per mg. Production of 1,25-(OH)2-D3 was linear with time up to 2 h and was dependent upon ferredoxin concentration as well as cytochrome P-450 concentration in the range of 0-40 nM. In the presence of excess ferredoxin and adequate amounts of cytochrome P-450, 1,25-(OH)2-D3 production was also dependent upon substrate concentrations in the range of 0.25 to 2.5 microM yielding an estimated Km of 1 microM. In the presence of excess substrate and ferredoxin, the catalytic-centre activity of the enzyme was estimated to be 1 h-1. | lld:pubmed |
| pubmed-article:2719667 | pubmed:grant | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:2719667 | pubmed:grant | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:2719667 | pubmed:grant | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:2719667 | pubmed:commentsCorrections | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:2719667 | pubmed:commentsCorrections | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:2719667 | pubmed:commentsCorrections | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:2719667 | pubmed:commentsCorrections | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:2719667 | pubmed:commentsCorrections | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:2719667 | pubmed:commentsCorrections | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:2719667 | pubmed:commentsCorrections | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:2719667 | pubmed:commentsCorrections | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:2719667 | pubmed:commentsCorrections | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:2719667 | pubmed:commentsCorrections | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:2719667 | pubmed:commentsCorrections | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:2719667 | pubmed:commentsCorrections | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:2719667 | pubmed:commentsCorrections | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:2719667 | pubmed:commentsCorrections | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:2719667 | pubmed:commentsCorrections | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:2719667 | pubmed:commentsCorrections | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:2719667 | pubmed:commentsCorrections | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:2719667 | pubmed:commentsCorrections | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:2719667 | pubmed:commentsCorrections | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:2719667 | pubmed:commentsCorrections | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:2719667 | pubmed:commentsCorrections | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:2719667 | pubmed:commentsCorrections | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:2719667 | pubmed:commentsCorrections | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:2719667 | pubmed:commentsCorrections | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:2719667 | pubmed:commentsCorrections | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:2719667 | pubmed:language | eng | lld:pubmed |
| pubmed-article:2719667 | pubmed:journal | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:2719667 | pubmed:citationSubset | IM | lld:pubmed |
| pubmed-article:2719667 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:2719667 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:2719667 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:2719667 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:2719667 | pubmed:chemical | http://linkedlifedata.com/r... | lld:pubmed |
| pubmed-article:2719667 | pubmed:status | MEDLINE | lld:pubmed |
| pubmed-article:2719667 | pubmed:month | Apr | lld:pubmed |
| pubmed-article:2719667 | pubmed:issn | 0264-6021 | lld:pubmed |
| pubmed-article:2719667 | pubmed:author | pubmed-author:GrayR WRW | lld:pubmed |
| pubmed-article:2719667 | pubmed:author | pubmed-author:GhazarianJ... | lld:pubmed |
| pubmed-article:2719667 | pubmed:issnType | Print | lld:pubmed |
| pubmed-article:2719667 | pubmed:day | 15 | lld:pubmed |
| pubmed-article:2719667 | pubmed:volume | 259 | lld:pubmed |
| pubmed-article:2719667 | pubmed:owner | NLM | lld:pubmed |
| pubmed-article:2719667 | pubmed:authorsComplete | Y | lld:pubmed |
| pubmed-article:2719667 | pubmed:pagination | 561-8 | lld:pubmed |
| pubmed-article:2719667 | pubmed:dateRevised | 2009-11-18 | lld:pubmed |
| pubmed-article:2719667 | pubmed:meshHeading | pubmed-meshheading:2719667-... | lld:pubmed |
| pubmed-article:2719667 | pubmed:meshHeading | pubmed-meshheading:2719667-... | lld:pubmed |
| pubmed-article:2719667 | pubmed:meshHeading | pubmed-meshheading:2719667-... | lld:pubmed |
| pubmed-article:2719667 | pubmed:meshHeading | pubmed-meshheading:2719667-... | lld:pubmed |
| pubmed-article:2719667 | pubmed:meshHeading | pubmed-meshheading:2719667-... | lld:pubmed |
| pubmed-article:2719667 | pubmed:meshHeading | pubmed-meshheading:2719667-... | lld:pubmed |
| pubmed-article:2719667 | pubmed:meshHeading | pubmed-meshheading:2719667-... | lld:pubmed |
| pubmed-article:2719667 | pubmed:meshHeading | pubmed-meshheading:2719667-... | lld:pubmed |
| pubmed-article:2719667 | pubmed:meshHeading | pubmed-meshheading:2719667-... | lld:pubmed |
| pubmed-article:2719667 | pubmed:meshHeading | pubmed-meshheading:2719667-... | lld:pubmed |
| pubmed-article:2719667 | pubmed:meshHeading | pubmed-meshheading:2719667-... | lld:pubmed |
| pubmed-article:2719667 | pubmed:year | 1989 | lld:pubmed |
| pubmed-article:2719667 | pubmed:articleTitle | Solubilization and reconstitution of kidney 25-hydroxyvitamin D3 1 alpha- and 24-hydroxylases from vitamin D-replete pigs. | lld:pubmed |
| pubmed-article:2719667 | pubmed:affiliation | Department of Medicine, Medical College of Wisconsin, Milwaukee 53226. | lld:pubmed |
| pubmed-article:2719667 | pubmed:publicationType | Journal Article | lld:pubmed |
| pubmed-article:2719667 | pubmed:publicationType | Research Support, U.S. Gov't, P.H.S. | lld:pubmed |
| http://linkedlifedata.com/r... | pubmed:referesTo | pubmed-article:2719667 | lld:pubmed |
| http://linkedlifedata.com/r... | pubmed:referesTo | pubmed-article:2719667 | lld:pubmed |
| http://linkedlifedata.com/r... | pubmed:referesTo | pubmed-article:2719667 | lld:pubmed |
| http://linkedlifedata.com/r... | pubmed:referesTo | pubmed-article:2719667 | lld:pubmed |
| http://linkedlifedata.com/r... | pubmed:referesTo | pubmed-article:2719667 | lld:pubmed |
