Elsevier

Biochimie Open

Volume 4, June 2017, Pages 99-106
Biochimie Open

Research paper
Subcellular localization of the five members of the human steroid 5α-reductase family

https://doi.org/10.1016/j.biopen.2017.03.003Get rights and content
Under a Creative Commons license
open access

Highlights

  • All members of human testosterone 5α-reductase family were expressed in HeLa cells.

  • Subcellular localization of SRD5A proteins in the endoplasmic reticulum is reported.

  • The effect of GFP tagging at N- or C-term on SRD5A proteins expression was assessed.

  • The TECRL gene is expressed for the first time and its product localizes in the ER.

Abstract

In humans the steroid 5α-reductase (SRD5A) family comprises five integral membrane enzymes that carry out reduction of a double bond in lipidic substrates: Δ4-3-keto steroids, polyprenol and trans-enoyl CoA. The best-characterized reaction is the conversion of testosterone into the more potent dihydrotestosterone carried out by SRD5A1-2. Some controversy exists on their possible nuclear or endoplasmic reticulum localization.

We report the cloning and transient expression in HeLa cells of the five members of the human steroid 5α-reductase family as both N- and C-terminus green fluorescent protein tagged protein constructs. Following the intrinsic fluorescence of the tag, we have determined that the subcellular localization of these enzymes is in the endoplasmic reticulum, upon expression in HeLa cells. The presence of the tag at either end of the polypeptide chain can affect protein expression and, in the case of trans enoyl-CoA reductase, it induces the formation of protein aggregates.

Keywords

Steroid 5α-reductase
Polyprenol reductase
Trans-enoyl-CoA reductase
Subcellular localization

Cited by (0)

1

These authors equally contributed to this work.