Evidence for distinct dehydrogenase and isomerase sites within a single 3.beta.-hydroxysteroid dehydrogenase/5-ene-4-ene isomerase protein
Abstract
Complementary DNA encoding human 3 beta-hydroxysteroid dehydrogenase/5-ene-4-ene isomerase (3 beta-HSD) has been expressed in transfected GH4C1 with use of the cytomegalovirus promoter. The activity of the expressed protein clearly shows that both dehydrogenase and isomerase enzymatic activities are present within a single protein. However, such findings do not indicate whether the two activities reside within one or two closely related catalytic sites. With use of [3H]-5-androstenedione, the intermediate compound in dehydroepiandrosterone (DHEA) transformation into 4-androstenedione by 3 beta-HSD, the present study shows that 4MA (N,N-diethyl-4-methyl-3-oxo-4-aza-5 alpha-androstane-17 beta-carboxamide) and its analogues inhibit DHEA oxidation competitively while they exert a noncompetitive inhibition of the isomerization of 5-androstenedione to 4-androstenedione with an approximately 1000-fold higher Ki value. The present results thus strongly suggest that dehydrogenase and isomerase activities are present at separate sites on the 3 beta-HSD protein. In addition, using 5 alpha-dihydrotestosterone (DHT) and 5 alpha-androstane-3 beta, 17 beta-diol as substrates for dehydrogenase activity only, we have found that dehydrogenase activity is reversibly and competitively inhibited by 4MA. Such data suggest that the irreversible step in the transformation of DHEA to 4-androstenedione is due to a separate site possessing isomerase activity that converts the 5-ene-3-keto to a much more stable 4-ene-3-keto configuration.
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HSD3B1 and HSD3B2 catalyze an irreversible reaction directly linked to isomerization of the Δ5 double bond.
“An exception to this rule are the two HSD3B isoforms, HSD3B1 and HSD3B2, which catalyze an irreversible reaction, directly linked to the isomerization of the Δ5 double bond.”
From Human steroid biosynthesis, metabolism and excretion are differentially reflected by serum and urine steroid metabolomes: a comprehensive review (2019) · cites this paper as [15]
HSD3B1 and HSD3B2 have dual catalytic activity: they transform the hydroxy group on carbon 3 to a keto group and isomerize the double bond from Δ5 to Δ4.
“These enzymes have dual catalytic activity and not only transform the hydroxy group on carbon 3 to a keto group but additionally isomerize the double bond from Δ5 to Δ4 [[15], [16], [17], [18]].”
From Human steroid biosynthesis, metabolism and excretion are differentially reflected by serum and urine steroid metabolomes: a comprehensive review (2019) · cites this paper as [15]
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- Authors
- Van, L. T., Takahashi, M., De Launoit, Y., Dumont, M., Lachance, Y., & Labrie, F.
- ORCID
- Dumont, Martine
- Venue
- Biochemistry
- Publisher
- American Chemical Society (ACS)
- Published
- 1991-09
- Volume
- 30
- Issue
- 36
- Pages
- 8861-8865
- ISSN
- 0006-2960 (ISSN-L)
- 1520-4995
- OpenAlex
- W2013149050
- Cited by
- 45
- References
- 0
- Citations by year
- 2023: 1 · 2019: 1 · 2018: 1 · 2015: 1 · 2013: 1 · 2012: 2
- FWCI
- 2.09
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Biochemistry (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Steroid Chemistry and Biochemistry
- Health Sciences › Medicine › Endocrinology, Diabetes and Metabolism › Adrenal Hormones and Disorders
- Health Sciences › Medicine › Endocrinology, Diabetes and Metabolism › Hormonal Regulation and Hypertension
- Keywords
- competitive inhibition, non-competitive inhibition, dehydrogenase activity, isomerase activity, dihydrotestosterone, 5α-androstane-3β,17β-diol, steroid metabolism
- MeSH terms
- Androgen Antagonists, Azasteroids, Binding Sites, Binding, Competitive, Cell Line, Dehydroepiandrosterone, Humans, Kinetics, Multienzyme Complexes, Placenta, Progesterone Reductase, Protein Conformation, Dihydrotestosterone, Steroid Isomerases, Substrate Specificity, Tumor Cells, Cultured, Androstane-3,17-diol
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