Cloning and production of antisera to human placental 11 β-hydroxysteroid dehydrogenase type 2
| Autor: | Robbie S. Lindsay, Caroline Mckenzie Leckie, Parvez Murad, John J. Mullins, Christopher R. W. Edwards, Yuri Kotelevtsev, Jonathan R. Seckl, Roger W. Brown, Lawrence P. Brett, Karen E. Chapman, Joyce L.W. Yau, Val Lyons |
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| Rok vydání: | 1996 |
| Předmět: |
Adult
Male medicine.medical_specialty DNA Complementary medicine.drug_class Placenta Molecular Sequence Data Carbenoxolone CHO Cells In situ hybridization Biology Transfection Polymerase Chain Reaction Biochemistry Antibodies chemistry.chemical_compound Fetus Pregnancy Corticosterone Cricetinae Internal medicine medicine Animals Humans Tissue Distribution Amino Acid Sequence RNA Messenger Cloning Molecular Molecular Biology In Situ Hybridization DNA Primers Aldosterone Base Sequence Molecular Structure Sequence Homology Amino Acid Hydroxysteroid Dehydrogenases Trophoblast Cell Biology Endocrinology medicine.anatomical_structure chemistry Mineralocorticoid 11-beta-Hydroxysteroid Dehydrogenases Female Rabbits Glucocorticoid Research Article medicine.drug |
| Zdroj: | Biochemical Journal. 313:1007-1017 |
| ISSN: | 1470-8728 0264-6021 |
| Popis: | By inactivating potent glucocorticoid hormones (cortisol and corticosterone), 11 beta-hydroxysteroid dehydrogenase type 2 (11 beta-HSD2) plays an important role in the placenta by controlling fetal exposure to maternal glucocorticoids, and in aldosterone target tissues by controlling ligand access to co-localized glucocorticoid and mineralocorticoid receptors. Amino acid sequence from homogeneous human placental 11 beta-HSD2 was used to isolate a 1897 bp cDNA encoding this enzyme (predicted M(r) 44126; predicted pI 9.9). Transfection into mammalian (CHO) cells produces 11 beta-HSD2 activity which is NAD(+)-dependent, is without reductase activity, avidly metabolizes glucocorticoids (Km values for corticosterone, cortisol and dexamethasone of 12.4 +/- 1.5, 43.9 +/- 8.5 and 119 +/- 15 nM respectively) and is inhibited by glycyrrhetinic acid and carbenoxolone (IC50 values 10-20 nM). Rabbit antisera recognizing 11 beta-HSD2 have been raised to an 11 beta-HSD2-(370--383)-peptide-carrier conjugate. Recombinant 11 beta-HSD2, like native human placental 11 beta-HSD2, is detectable with affinity labelling and anti-11 beta-HSD2 antisera, and appears to require little post-translational processing for activity. 11 beta-HSD2 mRNA (approximately 1.9 kb transcript) is expressed in placenta, aldosterone target tissues (kidney, parotid, colon and skin) and pancreas. In situ hybridization and immunohistochemistry localize abundant 11 beta-HSD2 expression to the distal nephron in human adult kidney and to the trophoblast in the placenta. 11 beta-HSD2 transcripts are expressed in fetal kidney (but not lung, liver or brain) at 21-26 weeks, suggesting that an 11 beta-HSD2 distribution resembling that in the adult is established by this stage in human development. |
| Databáze: | OpenAIRE |
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