Ban Nobuhiro, Sasaki Mayumi, Sakai Hiromichi, Ueda Kazumitsu, Inagaki Nobuya
Department of Physiology, Akita University School of Medicine, CREST of Japan Science and Technology Cooperation, 1-1-1, Hondo, Akita 010-8543, Japan.
Biochem J. 2005 Jul 15;389(Pt 2):577-85. doi: 10.1042/BJ20050159.
The A subclass of the ABC (ATP-binding cassette) transporter superfamily has a structural feature that distinguishes it from other ABC transporters, and is proposed to be involved in the transmembrane transport of endogenous lipids. Here we have cloned mouse and rat full-length cDNAs of ABCA17, a novel ABC transporter belonging to the A subclass. Mouse and rat ABCA17 proteins comprise 1733 and 1773 amino acid residues respectively, having 87.3% amino acid identity; mouse ABCA17 has amino acid identities of 55.3% and 36.7% with mouse ABCA3 and sea urchin ABCA respectively. RNA blot and quantitative real-time PCR analyses showed that ABCA17 mRNA is expressed exclusively in the testis. Examination of testis by in situ hybridization showed that ABCA17 mRNA is expressed in germ cells, mainly spermatocytes, in the seminiferous tubule. Immunoblot analysis using a specific antibody showed that ABCA17 is a protein of 200 kDa, and immunohistochemical analysis demonstrated that the protein is detected in the anterior head of sperm and elongated spermatids. ABCA17 was localized in the endoplasmic reticulum in transiently transfected HEK293 cells. Metabolic labelling analysis showed that intracellular esterified lipids, including cholesteryl esters, fatty acid esters and triacylglycerols, were significantly decreased in HEK293 cells stably expressing ABCA17 compared with untransfected cells. These results suggest that ABCA17 may play a role in regulating lipid composition in sperm.
ABC(ATP结合盒)转运蛋白超家族的A亚类具有使其有别于其他ABC转运蛋白的结构特征,据推测其参与内源性脂质的跨膜转运。在此,我们克隆了ABCA17的小鼠和大鼠全长cDNA,ABCA17是属于A亚类的一种新型ABC转运蛋白。小鼠和大鼠的ABCA17蛋白分别由1733和1773个氨基酸残基组成,氨基酸同一性为87.3%;小鼠ABCA17与小鼠ABCA3和海胆ABCA的氨基酸同一性分别为55.3%和36.7%。RNA印迹和定量实时PCR分析表明,ABCA17 mRNA仅在睾丸中表达。通过原位杂交对睾丸进行检测表明,ABCA17 mRNA在生精小管的生殖细胞中表达,主要是精母细胞。使用特异性抗体进行的免疫印迹分析表明,ABCA17是一种200 kDa的蛋白质,免疫组织化学分析表明,该蛋白质在精子头部前端和伸长的精子细胞中被检测到。ABCA17在瞬时转染的HEK293细胞中定位于内质网。代谢标记分析表明,与未转染的细胞相比,稳定表达ABCA17的HEK293细胞中包括胆固醇酯、脂肪酸酯和三酰甘油在内的细胞内酯化脂质显著减少。这些结果表明,ABCA17可能在调节精子脂质组成中发挥作用。