Differential activity and expression of human 5β-reductase (AKR1D1) splice variants

Steroid hormones, including glucocorticoids and androgens, exert a wide variety of effects in the body across almost all tissues. The steroid A-ring 5β-reductase (AKR1D1) is expressed in human liver and testes, and three splice variants have been identified (AKR1D1-001, AKR1D1-002, AKR1D1-006). Amongst these, AKR1D1-002 is the best described; it modulates steroid hormone availability and catalyses an important step in bile acid biosynthesis. However, specific activity and expression of AKR1D1-001 and AKR1D1-006 are unknown. Expression of AKR1D1 variants were measured in human liver biopsies and hepatoma cell lines by qPCR. Their three-dimensional (3D) structures were predicted using in silico approaches. AKR1D1 variants were overexpressed in HEK293 cells, and successful overexpression confirmed by qPCR and Western blotting. Cells were treated with either cortisol, dexamethasone, prednisolone, testosterone or androstenedione, and steroid hormone clearance was measured by mass spectrometry. Glucocorticoid and androgen receptor activation were determined by luciferase reporter assays. AKR1D1-002 and AKR1D1-001 are expressed in human liver, and only AKR1D1-006 is expressed in human testes. Following overexpression, AKR1D1-001 and AKR1D1-006 protein levels were lower than AKR1D1-002, but significantly increased following treatment with the proteasomal inhibitor, MG-132. AKR1D1-002 efficiently metabolised glucocorticoids and androgens and decreased receptor activation. AKR1D1-001 and AKR1D1-006 poorly metabolised dexamethasone, but neither protein metabolised cortisol, prednisolone, testosterone or androstenedione. We have demonstrated the differential expression and role of AKR1D1 variants in steroid hormone clearance and receptor activation in vitro. AKR1D1-002 is the predominant functional protein in steroidogenic and metabolic tissues. In addition, AKR1D1-001 and AKR1D1-006 may have a limited, steroid-specific role in the regulation of dexamethasone action.

Medienart:

E-Artikel

Erscheinungsjahr:

2021

Erschienen:

2021

Enthalten in:

Zur Gesamtaufnahme - volume:66

Enthalten in:

Journal of molecular endocrinology - 66(2021), 3 vom: 27. März, Seite 181-194

Sprache:

Englisch

Beteiligte Personen:

Appanna, Nathan [VerfasserIn]
Gibson, Hylton [VerfasserIn]
Gangitano, Elena [VerfasserIn]
Dempster, Niall J [VerfasserIn]
Morris, Karen [VerfasserIn]
George, Sherly [VerfasserIn]
Arvaniti, Anastasia [VerfasserIn]
Gathercole, Laura L [VerfasserIn]
Keevil, Brian [VerfasserIn]
Penning, Trevor M [VerfasserIn]
Storbeck, Karl-Heinz [VerfasserIn]
Tomlinson, Jeremy W [VerfasserIn]
Nikolaou, Nikolaos [VerfasserIn]

Links:

Volltext

Themen:

3-oxo-5 beta-steroid delta 4-dehydrogenase
Androgens
Cortisol
Dexamethasone
EC 1.-
EC 1.3.99.6
EC 3.4.25.1
Glucocorticoids
Journal Article
Liver
Mutant Proteins
Oxidoreductases
Proteasome Endopeptidase Complex
RNA, Messenger
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Steroids
Testosterone

Anmerkungen:

Date Completed 10.12.2021

Date Revised 16.02.2022

published: Print

Citation Status MEDLINE

doi:

10.1530/JME-20-0160

funding:

Förderinstitution / Projekttitel:

PPN (Katalog-ID):

NLM320641406