Regulation of the serotonin transporter by interacting proteins
Regulation of the serotonin transporter by interacting proteins
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DOI：
10.1042/0300-5127:0290722
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发表时间：
2001-01-01
影响因子：
3.9
通讯作者：
Williams, C
中科院分区：
文献类型：
--
作者：
Haase, J;Killian, AM;Williams, C
5-羟色胺转运体（SERT）在5-羟色胺（5-HT）维持正常神经传递中起着关键作用。最近的证据表明，SERT和其他神经递质转运蛋白受到严格调控。蛋白激酶C的激活导致SERT介导的5-HT摄取减少，这是由于转运蛋白的内化。然而，到目前为止，很少有人知道的机制和蛋白质参与下调的转运。一种候选的SERT调节蛋白是SNARE（可溶性N-乙基马来酰亚胺敏感因子附着蛋白受体）蛋白，突触融合蛋白1A（Syn 1A），其最近已经涉及离子通道以及SERT相关的γ-氨基丁酸和甘氨酸转运蛋白的调节。使用5-HT摄取测定，共聚焦显微镜和谷胱甘肽S-转移酶（GST）下拉测定，我们表明，Syn 1A也与SERT相互作用，改变了转运蛋白的亚细胞定位，导致5-HT转运减少。此外，我们还利用酵母双杂交系统寻找与SERT胞质N端结构域相互作用的新的调节蛋白。通过对大鼠脑cDNA文库的筛选，我们鉴定了6个潜在的SERT结合蛋白。在这里，我们还介绍了这些蛋白质的生物学相关性和它们对5-羟色胺转运蛋白的调节的潜在作用的阐明进展。
The serotonin transporter (SERT) plays a critical role in the maintenance of normal neurotransmission by serotonin [5-hydroxytryptamine (5-HT)]. Recent evidence suggests that SERT and other neurotransmitter transporters are tightly regulated. Activation of protein kinase C results in a decrease in SERT-mediated 5-HT uptake, which is due to an internalization of the transporter. However, to date little is known about the mechanism and proteins involved in the down-regulation of the transporter. One candidate SERT-regulatory protein is the SNARE (soluble N-ethylmaleimide-sensitive factor-attachment protein receptor) protein, syntaxin 1A (Syn1A), which has recently been implicated in the regulation of ion channels as well as the SERT-related gamma -aminobutyric acid- and glycine-transporters. Using 5-HT uptake assays, confocal microscopy and glutathione S-transferase (GST) pull-down assays we showed that Syn1A also interacts with SERT and alters the subcellular localization of the transporter, resulting in a reduction of 5-HT transport. In addition, we have used the yeast two-hybrid system to search for novel regulatory proteins that interact with the cytoplasmic N-terminal domain of SERT. By screening rat brain cDNA library we have identified six potential SERT-binding proteins. Here we also present progress towards the elucidation of the biological relevance of these proteins and their potential role for the regulation of the serotonin transporter.