Linked Life Data

7947689

Source:http://linkedlifedata.com/resource/pubmed/id/7947689

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
43
pubmed:dateCreated
1994-12-1
pubmed:abstractText
The human 11 beta-hydroxysteroid dehydrogenase (h11 beta-HSD) inactivates the active corticosteroid cortisol to its inactive metabolite cortisone. We have developed transactivation analyses of the reporter chimeric gene mouse mammary tumor virus-chloramphenicol acetyltransferase (MMTV-CAT) to study the catalytic activity of h11 beta-HSD introduced by cotransfection into receptor and 11 beta-HSD deficient CV-1 cells. Assay of 11 beta-HSD expressed in CV-1 cells by cotransfection showed that the catalyzed dehydrogenation of cortisol to cortisone was 2-fold higher in the presence of NADP. The reductase activity was dependent on the coenzyme NADPH. The addition of increasing concentrations of the inhibitor carbenoxolone (CBX) in the incubates blocked the enzyme activity in a dose dependent fashion. In CV-1 cells cotransfected with expression vectors of either human glucocorticoid (hGR1-777) or mineralocorticoid (hMR1-984) and the reporter plasmid MMTV-CAT, dexamethasone (DEX), aldosterone (ALDO), cortisol, and corticosterone induction of CAT activity was dose dependent. Cotransfection of CV-1 cells transfected with 10 micrograms of 11 beta-HSD expression vector reduced the transactivation of MMTV-CAT by hGR or hMR in the presence of either cortisol or corticosterone to basal values. The concomitant addition of 100 nM cortisone and 1 microM NADPH to these transfectants elevated CAT activity. These data show that transactivation analyses can be used to study the 11 beta-HSD-catalyzed regulation of corticosteroid levels, which triggers physiological processes and in certain cases provides an alternative to animal experimentation.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Nov
pubmed:issn
0006-2960
pubmed:author
pubmed:issnType
Print
pubmed:day
1
pubmed:volume
33
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
12837-43
pubmed:dateRevised
2008-11-21
pubmed:meshHeading
pubmed-meshheading:7947689-11-beta-Hydroxysteroid Dehydrogenases, pubmed-meshheading:7947689-Carbenoxolone, pubmed-meshheading:7947689-Cell Line, pubmed-meshheading:7947689-Chloramphenicol O-Acetyltransferase, pubmed-meshheading:7947689-Cortisone, pubmed-meshheading:7947689-Gene Expression Regulation, pubmed-meshheading:7947689-Genes, Reporter, pubmed-meshheading:7947689-Humans, pubmed-meshheading:7947689-Hydrocortisone, pubmed-meshheading:7947689-Hydroxysteroid Dehydrogenases, pubmed-meshheading:7947689-Mammary Tumor Virus, Mouse, pubmed-meshheading:7947689-NADP, pubmed-meshheading:7947689-Receptors, Glucocorticoid, pubmed-meshheading:7947689-Receptors, Mineralocorticoid, pubmed-meshheading:7947689-Recombinant Fusion Proteins, pubmed-meshheading:7947689-Transcription, Genetic, pubmed-meshheading:7947689-Transcriptional Activation, pubmed-meshheading:7947689-Transfection
pubmed:year
1994
pubmed:articleTitle
Transcription activation of mouse mammary tumor virus-chloramphenicol acetyltransferase: a model to study the metabolism of cortisol.
pubmed:affiliation
Medical Research Council Group in Molecular Endocrinology, CHUL Research Center, Québec, Canada.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't