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17β-Hydroxysteroid dehydrogenases types 1 and 2: Enzymatic assays based on radiometric and mass-spectrometric detection.
Methods Enzymol. 689, 201-234 (2023)
The 17β-hydroxysteroid dehydrogenase type 1 (HSD17B1) has a key role in estrogen biosynthesis as it catalyzes the reduction of estrone to the most potent estrogen, estradiol. Estradiol has a high affinity for estrogen receptors and thus stimulates their transactivation, which leads to cell proliferation and numerous other effects. HSD17B2 catalyzes the oxidation of estradiol to the less potent estrone, thereby decreasing estrogen receptor activation, which results in reduction of estrogen-associated effects. HSD17B1 and HSD17B2 overexpressing E.coli homogenates or recombinant enzymes can be used for screening and development of drugs against various pathologies such as cancer, endometriosis or osteoporosis. Here we describe the preparation of HSD17B1 and HSD17B2 bacterial homogenates and purified recombinant HSD17B1 protein as enzyme sources as well as enzymatic assays based on radiometric and mass-spectrometric detection for enzyme characterization.
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Publication type
Article: Journal article
Document type
Scientific Article
Keywords
E. Coli Overexpression ; Hsd17b1 ; Hsd17b2 ; Kinetic Characterization ; Recombinant Protein; Human Estrogenic 17-beta-hydroxysteroid-dehydrogenase; Placental Estradiol-17-beta Dehydrogenase; Expression; Inhibitors; Derivatives; Kinetics; Enzymes; Cancer; Tissue
ISSN (print) / ISBN
0076-6879
e-ISSN
0076-6879
Journal
Methods in Enzymology
Quellenangaben
Volume: 689,
Pages: 201-234
Publisher
Elsevier
Publishing Place
525 B Street, Suite 1900, San Diego, Ca 92101-4495 Usa
Non-patent literature
Publications
Reviewing status
Peer reviewed
Grants
Slovenian Research Agency