克隆和功能表征小鼠钠离子依赖型有机阴离子转运体 Soat(Slc10a6)。

Cloning and functional characterization of the mouse sodium-dependent organic anion transporter Soat (Slc10a6).

机构信息

Institute of Pharmacology and Toxicology, Justus Liebig University of Giessen, 35392 Giessen, Germany.

出版信息

J Steroid Biochem Mol Biol. 2013 Nov;138:90-9. doi: 10.1016/j.jsbmb.2013.03.009. Epub 2013 Apr 3.

Abstract

The sodium-dependent organic anion transporter SOAT is a member of the Solute Carrier Family SLC10. In man, this carrier is predominantly expressed in the testis and has transport activity for sulfoconjugated steroid hormones. Here, we report on cloning, expression analysis and functional characterization of the mouse Soat (mSoat) and compare its characteristics with the human SOAT carrier. Quantitative mRNA expression analysis for mSoat in male mice revealed very high expression in lung and further high expression in testis and skin. Immunohistochemical studies showed expression of the mSoat protein in bronchial epithelial cells of the lung, in primary and secondary spermatocytes as well as round spermatids within the seminiferous tubules of the testis, in the epidermis of the skin, and in the urinary epithelium of the bladder. Stably transfected mSoat-HEK293 cells revealed sodium-dependent transport for dehydroepiandrosterone sulfate (DHEAS), estrone-3-sulfate, and pregnenolone sulfate (PREGS) with apparent Km values of 60.3μM, 2.1μM, and 2.5μM, respectively. In contrast to human SOAT, which has a preference for DHEAS as a substrate, mSoat exhibits the highest transport rate for PREGS, likely reflecting differences in the steroid pattern between both species. In conclusion, although certain differences between human SOAT and mSoat exist regarding quantitative gene expression in endocrine and non-endocrine tissues, as well as in the transport kinetics for steroid sulfates, in general, both can be regarded as homologous carriers.

摘要

钠依赖性有机阴离子转运体 SOAT 是溶质载体家族 SLC10 的成员。在人类中,这种载体主要在睾丸中表达,并且对硫酸结合的甾体激素具有转运活性。在这里,我们报告了小鼠 Soat(mSoat)的克隆、表达分析和功能特征,并将其与人类 SOAT 载体进行了比较。对雄性小鼠中 mSoat 的定量 mRNA 表达分析显示,其在肺中表达水平非常高,在睾丸和皮肤中进一步高表达。免疫组织化学研究显示,mSoat 蛋白在肺的支气管上皮细胞、睾丸的初级和次级精母细胞以及精小管内的圆形精子中表达,在皮肤的表皮中和膀胱的尿路上皮中表达。稳定转染的 mSoat-HEK293 细胞显示出对脱氢表雄酮硫酸酯(DHEAS)、雌酮-3-硫酸酯和孕烯醇酮硫酸酯(PREGS)的钠依赖性转运,其表观 Km 值分别为 60.3μM、2.1μM 和 2.5μM。与优先作为底物的人类 SOAT 不同,mSoat 对 PREGS 的转运速率最高,这可能反映了两种物种之间甾体模式的差异。总之,尽管人类 SOAT 和 mSoat 之间在激素和非激素组织中的定量基因表达以及对甾体硫酸盐的转运动力学方面存在某些差异,但总体而言,它们都可以被视为同源载体。

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