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

Biochimie open Pub Date : 2017-06-01 DOI:10.1016/j.biopen.2017.03.003

Antonella Scaglione , Linda Celeste Montemiglio , Giacomo Parisi , Italia Anna Asteriti , Renato Bruni , Gabriele Cerutti , Claudia Testi , Carmelinda Savino , Filippo Mancia , Patrizia Lavia , Beatrice Vallone

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引用次数: 11

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.

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人类固醇5α-还原酶家族5个成员的亚细胞定位

在人类中，类固醇5α-还原酶(SRD5A)家族包括五种完整的膜酶，它们在脂质底物中进行双键还原:Δ4-3-keto类固醇、聚戊二醇和反式烯丙醇辅酶a。最典型的反应是由SRD5A1-2将睾酮转化为更有效的双氢睾酮。它们在核或内质网的定位存在争议。我们克隆了5α-还原酶家族的5个成员，并在HeLa细胞中瞬时表达为N端和c端绿色荧光蛋白标记的蛋白构建体。根据标签的固有荧光，我们已经确定这些酶的亚细胞定位在内质网中，在HeLa细胞中表达。标签在多肽链两端的存在可以影响蛋白质表达，在反式烯酰辅酶a还原酶的情况下，它诱导蛋白质聚集体的形成。

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Biochimie open

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