Lee Woon Kyu, Hwang Ji-Sun, Yun Cheol-Heui, Cha Seok Ho
Department of Laboratory Animal Medicine Medical Research Center, College of Medicine, Yonsei University, Seoul 120-752, Korea.
Exp Mol Med. 2007 Dec 31;39(6):787-95. doi: 10.1038/emm.2007.85.
Organic ion transporters are expressed in various tissues that transport endogenous and exogenous compounds including their metabolites. There are organic anion transporter (OAT), organic cation transporter (OCT), organic anion transporter like protein (OATLP) and organic cation transporter like (OCTL). Considering the variety of charged organic ionic compounds, the existence of numerous isoforms of organic ion transporters can be assumed. In the present study, we have searched for a new isoform in the expressed sequence tag (EST) database using human organic anion transporter 4 (hOAT4) amino acid sequence as a "query". We found a candidate clone (BC021449) from the mouse kidney cDNA library. This clone was identified as an ortholog of ORCTL3 or OCTL-1. The mOCTL1 cDNA consists of 2016 base pairs encoding 551 amino acid residues with 12 putative transmembrane domains. The deduced amino acid sequence of mOCTL1 showed 35 to 40% identity to those of the other members of the OATs and OCTs. According to the tissue distribution, examined by Northern blot analysis, about a 2.4-kb transcript of mOCTL1 was observed in the kidney. About a 90-kDa band was detected when Western blot analysis in the mouse kidney was done by using antibody against synthesized oligopeptide of mOCTL1. The immunohistochemical result showed that mOCTL1 was stained at the glomerulus (the parietal epithelial cells and podocytes), pars recta of proximal tubule, distal convoluted tubules, connecting tubules and collecting tubules. From these results, we conclude that mOCTL1 may be a candidate for an organic ion transporter isoform in the mouse kidney.
有机离子转运体表达于多种组织中,可转运内源性和外源性化合物及其代谢产物。有机离子转运体包括有机阴离子转运体(OAT)、有机阳离子转运体(OCT)、有机阴离子转运体样蛋白(OATLP)和有机阳离子转运体样蛋白(OCTL)。鉴于带电有机离子化合物种类繁多,可以推测存在众多有机离子转运体亚型。在本研究中,我们以人类有机阴离子转运体4(hOAT4)氨基酸序列作为“查询序列”,在表达序列标签(EST)数据库中搜索新的亚型。我们从小鼠肾脏cDNA文库中发现了一个候选克隆(BC021449)。该克隆被鉴定为ORCTL3或OCTL-1的直系同源物。mOCTL1 cDNA由2016个碱基对组成,编码551个氨基酸残基,有12个推定的跨膜结构域。mOCTL1推导的氨基酸序列与OATs和OCTs其他成员的序列有35%至40%的同一性。根据Northern印迹分析检测的组织分布,在肾脏中观察到约2.4kb的mOCTL1转录本。当使用针对mOCTL1合成寡肽的抗体对小鼠肾脏进行Western印迹分析时,检测到约90kDa的条带。免疫组织化学结果显示,mOCTL1在肾小球(壁层上皮细胞和足细胞)、近端小管直部、远曲小管、连接小管和集合小管中被染色。从这些结果中,我们得出结论,mOCTL1可能是小鼠肾脏中有机离子转运体亚型的一个候选者。