Cloning and Expression of Rat cDNA Encoding Corticosteroid 11β-Dehydrogenase

Corticosteroid 11β-dehydrogenase (11-DH) catalyzes the conversion of cortisol to the inactive metabolite cortisone. Absence of 11-DH activity leads to a potentially fatal form of childhood hypertension termed apparent mineralocorticoid excess. As a first step in elucidating the molecular basis of th...

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Bibliographic Details
Published in:The Journal of biological chemistry 1989-11, Vol.264 (32), p.18939-18943
Main Authors: Agarwal, A K, Monder, C, Eckstein, B, White, P C
Format: Article
Language:English
Subjects:
11-beta-Hydroxysteroid Dehydrogenases
Amino Acid Sequence
Animals
Base Sequence
Biological and medical sciences
Cloning, Molecular
DNA - genetics
DNA - isolation & purification
Fundamental and applied biological sciences. Psychology
Gene Expression
Gene Library
Genes
Genes. Genome
Humans
Hydroxysteroid Dehydrogenases - genetics
Liver - enzymology
Molecular and cellular biology
Molecular genetics
Molecular Sequence Data
Nucleic Acid Hybridization
Rats
Restriction Mapping
Sequence Homology, Nucleic Acid
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Summary:Corticosteroid 11β-dehydrogenase (11-DH) catalyzes the conversion of cortisol to the inactive metabolite cortisone. Absence of 11-DH activity leads to a potentially fatal form of childhood hypertension termed apparent mineralocorticoid excess. As a first step in elucidating the molecular basis of this disorder, we isolated and characterized a rat cDNA clone encoding 11-DH. This clone hybridized to a single mRNA species in liver, kidney, and testis RNA but not to RNA from heart. The insert was 1265 base pairs long and included an 861-base pair open reading frame encoding 287 amino acids. A search of sequence databases revealed that 11-DH is identical in about 27% of amino acid residues to ribitol dehydrogenase from Klebsiella and to the product of the nodG gene from the nitrogen-fixing bacterium, Rhizobium meliloti, thus defining a new superfamily of genes encoding dehydrogenases. The 11-DH cDNA was expressed by transfection into Chinese hamster ovary cells under the control of an SV40 promoter. The expressed enzyme mediated both 11β-dehydrogenation and the reverse 11-oxoreduction reaction. Southern blot analysis of rat and human DNA suggested that additional genes related to 11-DH exist in both species.
ISSN:0021-9258
1083-351X
DOI:10.1016/S0021-9258(19)47248-7