Scaglione Antonella, Montemiglio Linda Celeste, Parisi Giacomo, Asteriti Italia Anna, Bruni Renato, Cerutti Gabriele, Testi Claudia, Savino Carmelinda, Mancia Filippo, Lavia Patrizia, Vallone Beatrice
Dept. of Biochemical Sciences, Sapienza University of Rome, P.le A.Moro 5, 00185, Rome, Italy.
Istituto Pasteur-Fondazione Cenci Bolognetti Dept. of Biochemical Sciences, Sapienza University of Rome.
Biochim Open. 2017 Jun;4:99-106. doi: 10.1016/j.biopen.2017.03.003. Epub 2017 Mar 21.
In humans the steroid 5alpha-reductase (SRD5A) family comprises five integral membrane enzymes that carry out reduction of a double bond in lipidic substrates: Δ-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.
在人类中，类固醇5α-还原酶（SRD5A）家族由五种整合膜酶组成，这些酶可对脂质底物中的双键进行还原反应：Δ-3-酮类固醇、聚戊烯醇和反式烯酰辅酶A。研究最为透彻的反应是由SRD5A1-2将睾酮转化为活性更强的双氢睾酮。关于它们可能定位于细胞核还是内质网存在一些争议。我们报告了人类类固醇5α-还原酶家族五个成员作为N端和C端均带有绿色荧光蛋白标记的蛋白质构建体在HeLa细胞中的克隆及瞬时表达。通过标签的固有荧光，我们确定在HeLa细胞中表达时，这些酶的亚细胞定位在内质网。多肽链两端存在标签会影响蛋白质表达，对于反式烯酰辅酶A还原酶而言，它会诱导蛋白质聚集体的形成。
