小鼠Rdh13基因的组织表达谱及其亚细胞定位的初步观察

目的：研究视黄醇脱氢酶13（Rdh13）基因在小鼠组织中的表达谱及其亚细胞定位,为其功能研究提供线索。方法：利用生物信息学方法模拟Rdh13的三维结构,采用半定量RT-PCR和Western blot的方法检测小鼠14种组织中Rdh13的表达水平;采用Western blot的方法检测其亚细胞分布;进一步应用免疫荧光共定位的方法观察Rdh13的亚细胞定位。结果：Rdh13具有SDR家族保守的辅酶结合位点（TGX3GXG）和催化活性位点（YX3K）;RT-PCR和Western blot实验证实Rdh13在小鼠多种组织中广泛表达,但其表达水平存在一定差异;不同于其它RDHs家族成员,Western blot和免疫荧光共定位提示Rdh13蛋白在细胞中定位于线粒体。结论：Rdh13是一个与Rdh12结构相似的SDR家族成员,在小鼠多个组织中广泛表达;Rdh13蛋白定位于线粒体,有可能作为保护线粒体不受视黄醛毒性作用的屏障。

A preliminary study on the expression profile of murine dehydrogenase 13 （Rdhl 3） and its subcellular localization

Objective： To study the expression profile ofretinol dehydrogenase 13 （Rdh13） in mouse tissues and determine its subcellular localization. Methods： Bioinformatics analysis was used to predict tertiary structure of Rdh13. The gene expression of Rdh13 in 14 tissues of mouse was examined by semi-quantitative RT-PCR and Western blot. The subcellular localization of Rdh13 was detected by using Western blot and confirmed by immunofluorescence staining. Results： Rdh13, like other SDRs, also harbors the highly conserved domain with a coenzyme-binding motif （TGX3GXG） and a catalytically active site （NX3K）. RT-PCR and Western blot show that Rdh13 ubiquitously expresses in various mouse tissues with different expression level. Western blot and immunofluorescence staining reveal that Rdh13, unlike other members of RDH family, is localized in mitochondria. Conclusions： Rdh 13, a member of SDR superfamily, shares similar tertiary structure with RDH 12. It is widely expressed in mouse tissues and localized in mitochondria, suggesting that Rdh 13 may serve as a barrier protecting the mitochondria against the highly reactive retinaldehyde.