Dutta B, Huang W, Molero M, Kekuda R, Leibach F H, Devoe L D, Ganapathy V, Prasad P D
Department of Obstetrics, Medical College of Georgia, Augusta, Georgia 30912, USA.
J Biol Chem. 1999 Nov 5;274(45):31925-9. doi: 10.1074/jbc.274.45.31925.
We have isolated a cDNA from human placenta, which, when expressed heterologously in mammalian cells, mediates the transport of the water-soluble vitamin thiamine. The cDNA codes for a protein of 497 amino acids containing 12 putative transmembrane domains. Northern blot analysis indicates that this transporter is widely expressed in human tissues. When expressed in HeLa cells, the cDNA induces the transport of thiamine (K(t) = 2.5 +/- 0.6 microM) in a Na(+)-independent manner. The cDNA-mediated transport of thiamine is stimulated by an outwardly directed H(+) gradient. Substrate specificity assays indicate that the transporter is specific to thiamine. Even though thiamine is an organic cation, the cDNA-induced thiamine transport is not inhibited by other organic cations. Similarly, thiamine is not a substrate for the known members of mammalian organic cation transporter family. The thiamine transporter gene, located on human chromosome 1q24, consists of 6 exons and is most likely the gene defective in the metabolic disorder, thiamine-responsive megaloblastic anemia. At the level of amino acid sequence, the thiamine transporter is most closely related to the reduced-folate transporter and thus represents the second member of the folate transporter family.
我们从人胎盘中分离出一种互补DNA(cDNA),当它在哺乳动物细胞中进行异源表达时,可介导水溶性维生素硫胺素的转运。该cDNA编码一种含有497个氨基酸的蛋白质,其中有12个推定的跨膜结构域。Northern印迹分析表明,这种转运蛋白在人体组织中广泛表达。当在HeLa细胞中表达时,该cDNA以不依赖钠离子的方式诱导硫胺素的转运(转运常数K(t)=2.5±0.6微摩尔)。cDNA介导的硫胺素转运受到外向性氢离子梯度的刺激。底物特异性分析表明,该转运蛋白对硫胺素具有特异性。尽管硫胺素是一种有机阳离子,但cDNA诱导的硫胺素转运不受其他有机阳离子的抑制。同样,硫胺素也不是哺乳动物有机阳离子转运蛋白家族已知成员的底物。硫胺素转运蛋白基因位于人类1号染色体的1q24区域,由6个外显子组成,很可能是导致代谢紊乱——硫胺素反应性巨幼细胞贫血的缺陷基因。在氨基酸序列水平上,硫胺素转运蛋白与还原型叶酸转运蛋白关系最为密切,因此它是叶酸转运蛋白家族的第二个成员。