Molecular Cloning of Testicular 20α-Hydroxysteroid Dehydrogenase

Identity with Aldose Reductase

James C. Warren, Gary L. Murdock, Yupo Ma, Steven Goodman, Warren E. Zimmer

Research output: Contribution to journalArticle

81 Citations (Scopus)

Abstract

Complementary DNA (cDNA) clones encoding bovine testicular 20α-hydroxysteroid dehydrogenase (20α-HSD) have been isolated from a bovine testicular λgt11 library using polyclonal antibodies against 20α-HSD and DNA probe hybridization. Nucleotide sequencing of three independently isolated clones was used to establish a composite cDNA sequence that encodes the enzyme. It contains a coding sequence of 921 nucleotides, a stop codon, and a 264-nucleotide 3′-noncoding segment which allowed deduction of the amino acid sequence of the enzyme. A computer homology search of the 20α-HSD cDNA performed against the GenBank DNA sequence database revealed it to be identical with bovine lens aldose reductase (alditol:NADPH oxidoreductase; EC 1.1.1.21), and a literature search reveals the deduced amino acid sequence to be identical with that reported for the bovine enzyme. Sequences obtained from the N-terminus of purified testicular 20α-HSD and from random peptides obtained by treatment with endopeptidase Lys-C are all identical with regions of the deduced amino acid sequence of 20α-HSD and/or the published sequence of aldose reductase. Further, the enzyme purified to homogeneity by following activity with 17-hydroxyprogesterone as a substrate was shown to reduce glucose, glyceraldehyde, and benzaldehyde (all classic aldose reductase substrates). Finally, 17-hydroxyprogesterone inhibited the reduction of benzaldehyde and glyceraldehyde. Because aldose reductase has been implicated in the etiology of diabetic complications, acceptance of steroid substrates may offer new implications for therapy.

Original languageEnglish (US)
Pages (from-to)1401-1406
Number of pages6
JournalBiochemistry
Volume32
Issue number6
DOIs
StatePublished - Jan 1 1993

Fingerprint

20-Hydroxysteroid Dehydrogenases
Aldehyde Reductase
Cloning
Molecular Cloning
Glyceraldehyde
17-alpha-Hydroxyprogesterone
Amino Acid Sequence
Nucleotides
Complementary DNA
DNA sequences
Nucleic Acid Databases
Enzymes
Amino Acids
Substrates
Clone Cells
Sugar Alcohols
Terminator Codon
DNA Probes
Diabetes Complications
NADP

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Molecular Cloning of Testicular 20α-Hydroxysteroid Dehydrogenase : Identity with Aldose Reductase. / Warren, James C.; Murdock, Gary L.; Ma, Yupo; Goodman, Steven; Zimmer, Warren E.

In: Biochemistry, Vol. 32, No. 6, 01.01.1993, p. 1401-1406.

Research output: Contribution to journalArticle

Warren, James C. ; Murdock, Gary L. ; Ma, Yupo ; Goodman, Steven ; Zimmer, Warren E. / Molecular Cloning of Testicular 20α-Hydroxysteroid Dehydrogenase : Identity with Aldose Reductase. In: Biochemistry. 1993 ; Vol. 32, No. 6. pp. 1401-1406.
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abstract = "Complementary DNA (cDNA) clones encoding bovine testicular 20α-hydroxysteroid dehydrogenase (20α-HSD) have been isolated from a bovine testicular λgt11 library using polyclonal antibodies against 20α-HSD and DNA probe hybridization. Nucleotide sequencing of three independently isolated clones was used to establish a composite cDNA sequence that encodes the enzyme. It contains a coding sequence of 921 nucleotides, a stop codon, and a 264-nucleotide 3′-noncoding segment which allowed deduction of the amino acid sequence of the enzyme. A computer homology search of the 20α-HSD cDNA performed against the GenBank DNA sequence database revealed it to be identical with bovine lens aldose reductase (alditol:NADPH oxidoreductase; EC 1.1.1.21), and a literature search reveals the deduced amino acid sequence to be identical with that reported for the bovine enzyme. Sequences obtained from the N-terminus of purified testicular 20α-HSD and from random peptides obtained by treatment with endopeptidase Lys-C are all identical with regions of the deduced amino acid sequence of 20α-HSD and/or the published sequence of aldose reductase. Further, the enzyme purified to homogeneity by following activity with 17-hydroxyprogesterone as a substrate was shown to reduce glucose, glyceraldehyde, and benzaldehyde (all classic aldose reductase substrates). Finally, 17-hydroxyprogesterone inhibited the reduction of benzaldehyde and glyceraldehyde. Because aldose reductase has been implicated in the etiology of diabetic complications, acceptance of steroid substrates may offer new implications for therapy.",
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