Molecular cloning and expression of rat liver 3α-hydroxysteroid dehydrogenase

Kuo Chi Cheng, Perrin C. White, Ke Nan Qin

Research output: Contribution to journalArticlepeer-review

67 Scopus citations

Abstract

Complementary DNA clones encoding 3α-hydroxysteroid dehydrogenase (SaHSD) were isolated from a rat liver cDNA λgt11 expression library using monoclonal antibodies as probes. The sizes of the cDNA inserts ranged from 1.3-2.3 kilobases. Sequence analysis indicated that variation in the DNA size was due to heterogeneity in the length of 3′ noncoding sequences. A full-length cDNA clone of 1286 basepairs contained an open reading frame encoding a protein of 322 amino acids with an estimated mol wt of 37 KDa. When expressed in E. coli, the encoded protein migrated to the same position on sodium dodecyl sulfate-polyacrylamide gels as the enzyme purified from rat liver cytosols. The protein expressed in bacteria was highly active in androsterone reduction in the presence of NAD as cofactor, and this activity was inhibited by indomethacin, a potent inhibitor of 3aHSD. The predicted amino acid sequence of 3aHSD was related to sequences of several other enzymes, including bovine prostaglandin F synthase, human chlordecone reductase, human aldose reductase, human aldehyde reductase, and frog lens ∈-crystalline, suggesting that these proteins belong to the same gene family.

Original languageEnglish (US)
Pages (from-to)823-828
Number of pages6
JournalMolecular Endocrinology
Volume5
Issue number6
StatePublished - Jun 1991

ASJC Scopus subject areas

  • Molecular Biology
  • Endocrinology

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