Ugawa S, Sunouchi Y, Ueda T, Takahashi E, Saishin Y, Shimada S
Department of Anatomy II, Nagoya City University Medical School, 1 Kawasumi, Mizuho-cho, Mizuho-ku, Nagoya 467-8601, Japan.
Am J Physiol Gastrointest Liver Physiol. 2001 Aug;281(2):G365-70. doi: 10.1152/ajpgi.2001.281.2.G365.
Previous experiments have shown that an amino acid transport system B(0+) transporter in cultured colonic epithelial cells mediates amino acid absorption. Here we describe the cloning and functional characterization of a system B(0+) transporter selectively expressed in the colon. Using the combination of an expressed sequence tag database search and RT-PCR approaches, we cloned a mouse colonic amino acid transporter, designated mCATB(0+). Northern blot analysis revealed that mCATB(0+) was selectively expressed in the large intestine. In situ hybridization showed the mCATB(0+) mRNA to be localized in absorptive epithelial cells. When expressed in Xenopus oocytes, mCATB(0+) exhibited a Na(+)-dependent stereoselective uptake and a broad specificity for neutral and cationic amino acids, which is characteristic of amino acid transport system B(0+). In vivo [(3)H]glycine uptake assay demonstrated that a system B(0+)-like transporter protein was expressed on the apical surface of the colonic absorptive cells. Our data suggest that a mouse colonic amino acid transporter mCATB(0+) may absorb amino acids from the intestinal contents in the colon.
先前的实验表明,培养的结肠上皮细胞中的一种氨基酸转运系统B(0+)转运体介导氨基酸吸收。在此,我们描述了一种在结肠中选择性表达的系统B(0+)转运体的克隆及功能特性。利用表达序列标签数据库搜索和逆转录聚合酶链反应(RT-PCR)方法相结合,我们克隆了一种小鼠结肠氨基酸转运体,命名为mCATB(0+)。Northern印迹分析显示,mCATB(0+)在大肠中选择性表达。原位杂交表明,mCATB(0+) mRNA定位于吸收性上皮细胞中。当在非洲爪蟾卵母细胞中表达时,mCATB(0+)表现出钠依赖性立体选择性摄取以及对中性和阳离子氨基酸的广泛特异性,这是氨基酸转运系统B(0+)的特征。体内[³H]甘氨酸摄取试验表明,一种类似系统B(0+)的转运体蛋白表达于结肠吸收细胞的顶端表面。我们的数据表明,小鼠结肠氨基酸转运体mCATB(0+)可能从结肠中的肠内容物吸收氨基酸。