Cloning, characterization, and expression analysis of a putative 17 beta-hydroxysteroid dehydrogenase 11 in the abalone, Haliotis diversicolor supertexta

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• 作者：Hong-ning Zhai^a ; ^b ; Jin Zhou^b ; Zhong-hua Cai^b ; ^{caizh@sz.tsinghua.edu.cn}
• 关键词：17尾-HSD-11 ; 17尾-hydroxysteroid dehydrogenase 11 ; SDRs ; short-chain dehydrogenases/reductases ; E1 ; estrone ; E2 ; estradiol ; 3伪-diol ; 5伪-androstane-3伪 ; 17尾-diol ; A ; androsterone ; T ; testosterone ; 4A ; androstenedione ; DMSO ; dimethyl sulfoxide ; ORF ; open
• 刊名：The Journal of Steroid Biochemistry and Molecular Biology
• 出版年：2012
• 期刊代码：172_09600760
• 类别：med
• 出版时间：May, 2012
• 卷：130
• 期：1-2
• 页码：57-63
• 文件大小：855 K

摘要

The 17-beta-hydroxysteroid dehydrogenases (17尾-HSDs) are key enzymes for sex steroid biosynthesis. To date, relatively little is known about the presence and function of 17尾-HSDs in marine gastropods. In the present study, a cDNA sequence encoding putative 17尾-HSD type 11 (17尾-HSD-11) was identified in marine abalone (Haliotis diversicolor supertexta). The full-length cDNA contains 1058 bp, including an open reading frame (ORF) of 900 bp that encodes a protein of 299 amino acids. Comparative structural analysis revealed that abalone 17尾-HSD-11 shares relatively high homology with other 17b-HSD-11 hormologues, and a lesser degree of amino acid identity with other forms of 17b-HSD, especially in the functional domains, including the cofactor binding domain (TGxxxGxG) and catalytic site (YxxSK). Phylogenetic analysis showed that abalone 17尾-HSD-11 belongs to the short-chain dehydrogenase/reductase (SDR) family. Functional analysis following transient transfection of the ORF into human embryonic kidney-293 (HEK-293) cells indicated that abalone 17尾-HSD-11 has the ability to convert 5伪-androstane-3伪,17尾-diol (3伪-diol) to androsterone (A) and testosterone (T) to androstenedione (4A). Expression analysis in vivo demonstrated that abalone 17尾-HSD-11 is differentially expressed during three stages (non-reproductive, reproductive, and post-reproductive). Taken together, these results indicate that ab-17尾-HSD-11 is an SDR family member with a potential role in steroid regulation during the reproductive stage.