In humans the steroid 5a-reductase (SRD5A) family comprises five integral membrane enzymes that carry out reduction of a double bond in lipidic substrates: D4-3-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 5a-reductase family as both N- and Cterminus 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.
Subcellular localization of the five members of the human steroid 5α-reductase family / Scaglione, Antonella; Montemiglio, LINDA CELESTE; Parisi, Giacomo; Asteriti, ITALIA ANNA; Bruni, Renato; Cerutti, Gabriele; Testi, Claudia; Savino, Carmelinda; Mancia, Filippo; Lavia, Patrizia; Vallone, Beatrice. - In: BIOCHIMIE OPEN. - ISSN 2214-0085. - ELETTRONICO. - 4:(2017), pp. 99-106. [10.1016/j.biopen.2017.03.003]
Subcellular localization of the five members of the human steroid 5α-reductase family
SCAGLIONE, ANTONELLA;MONTEMIGLIO, LINDA CELESTE;PARISI, GIACOMO;ASTERITI, ITALIA ANNA;CERUTTI, GABRIELE;TESTI, CLAUDIA;VALLONE, Beatrice
2017
Abstract
In humans the steroid 5a-reductase (SRD5A) family comprises five integral membrane enzymes that carry out reduction of a double bond in lipidic substrates: D4-3-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 5a-reductase family as both N- and Cterminus 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.File | Dimensione | Formato | |
---|---|---|---|
Scaglione_Subcellular_2017
accesso aperto
Note: Articolo principale
Tipologia:
Documento in Post-print (versione successiva alla peer review e accettata per la pubblicazione)
Licenza:
Creative commons
Dimensione
1.84 MB
Formato
Adobe PDF
|
1.84 MB | Adobe PDF |
I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.