Brüss M, Pörzgen P, Bryan-Lluka L J, Bönisch H
Institute of Pharmacology and Toxicology, University of Bonn, Germany.
Brain Res Mol Brain Res. 1997 Dec 15;52(2):257-62. doi: 10.1016/s0169-328x(97)00267-2.
The rat norepinephrine transporter (rNET) cDNA from the PC12 pheochromocytoma cell line has been cloned by RT-PCR and characterized. The cDNA encodes an integral membrane protein consisting of 617 amino acids which contains twelve putative transmembrane domains, two potential N-glycosylation sites, two potential phosphorylation sites for protein kinase C and one phosphorylation site for casein kinase II. The nucleotide and deduced amino acid sequence shows a high level of homology to the human and the bovine norepinephrine transporter and less homology to the rat dopamine transporter (rDAT). Heterologous expression of rNET in HEK293 cells revealed that uptake of [3H]norepinephrine is sodium- and chloride-dependent and highly sensitive to the selective norepinephrine transporter inhibitors desipramine and nisoxetine. The cloned rNET cDNA provides the opportunity to investigate this transporter in heterologous expression systems and adds a new member to the family of sodium- and chloride-dependent neurotransmitter transporters.
通过逆转录聚合酶链反应（RT-PCR）克隆并鉴定了来自PC12嗜铬细胞瘤细胞系的大鼠去甲肾上腺素转运体（rNET）cDNA。该cDNA编码一种由617个氨基酸组成的整合膜蛋白，包含12个推定的跨膜结构域、2个潜在的N-糖基化位点、2个蛋白激酶C的潜在磷酸化位点和1个酪蛋白激酶II的磷酸化位点。核苷酸和推导的氨基酸序列与人和牛的去甲肾上腺素转运体具有高度同源性，与大鼠多巴胺转运体（rDAT）的同源性较低。rNET在HEK293细胞中的异源表达表明，[3H]去甲肾上腺素的摄取依赖于钠和氯，并且对选择性去甲肾上腺素转运体抑制剂地昔帕明和尼索西汀高度敏感。克隆的rNET cDNA为在异源表达系统中研究该转运体提供了机会，并为钠和氯依赖性神经递质转运体家族增添了一个新成员。
