Allosteric modulatory effects of SRI-20041 and SRI-30827 on cocaine and HIV-1 Tat protein binding to human dopamine transporter.
Allosteric modulatory effects of SRI-20041 and SRI-30827 on cocaine and HIV-1 Tat protein binding to human dopamine transporter.
复制标题
SRI-20041 和 SRI-30827 对可卡因和 HIV-1 Tat 蛋白与人多巴胺转运蛋白结合的变构调节作用。
DOI：
10.1038/s41598-017-03771-0
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发表时间：
2017
影响因子：
4.6
通讯作者：
Zhu,Jun
中科院分区：
文献类型：
--
作者：
Sun,Wei-Lun;Quizon,PamelaM;Yuan,Yaxia;Zhang,Wei;Ananthan,Subramaniam;Zhan,Chang-Guo;Zhu,Jun
多巴胺转运蛋白（DAT）是可卡因和HIV-1转录反式激活因子（达特）蛋白的作用靶点。鉴定具有可卡因和达特与DAT结合的潜在衰减的变构调节分子具有很大的科学和临床意义。我们证明，酪氨酸470和88作为功能识别残基在人DAT（hDAT）的Tat诱导的抑制DA运输和转运蛋白构象转换。在这里，我们研究了两种变构配体SRI-20041和SRI-30827对可卡因结合野生型（WT）hDAT，Y 470 H和Y88 F突变体的变构调节作用。还测定了SRI-30827对Tat诱导的[3 H] WIN 35，428结合抑制的影响。与竞争性DAT抑制剂茚达曲林相比，两种SRI化合物在WT hDAT中抑制可卡因摄取[3 H]DA的IC 50中显示出相似的降低（30%）。相对于单独的可卡因，在可卡因之后添加SRI-20041或SRI-30827减慢了WT hDAT中[3 H] WIN 35，428结合的解离速率。此外，Y 470 H和Y88 F hDAT增强可卡因对DA摄取的抑制作用，并减弱SRI化合物对可卡因介导的解离速率的影响。SRI-30827减弱了Tat诱导的[3 H] WIN 35，428结合抑制。这些观察结果表明，酪氨酸470和88对于SRI化合物对可卡因与hDAT相互作用的变构调节作用是关键的。
Dopamine transporter (DAT) is the target of cocaine and HIV-1 transactivator of transcription (Tat) protein. Identifying allosteric modulatory molecules with potential attenuation of cocaine and Tat binding to DAT are of great scientific and clinical interest. We demonstrated that tyrosine 470 and 88 act as functional recognition residues in human DAT (hDAT) for Tat-induced inhibition of DA transport and transporter conformational transitions. Here we investigated the allosteric modulatory effects of two allosteric ligands, SRI-20041 and SRI-30827 on cocaine binding on wild type (WT) hDAT, Y470 H and Y88 F mutants. Effect of SRI-30827 on Tat-induced inhibition of [3H]WIN35,428 binding was also determined. Compared to a competitive DAT inhibitor indatraline, both SRI-compounds displayed a similar decrease (30%) in IC50for inhibition of [3H]DA uptake by cocaine in WT hDAT. The addition of SRI-20041 or SRI-30827 following cocaine slowed the dissociation rate of [3H]WIN35,428 binding in WT hDAT relative to cocaine alone. Moreover, Y470H and Y88F hDAT potentiate the inhibitory effect of cocaine on DA uptake and attenuate the effects of SRI-compounds on cocaine-mediated dissociation rate. SRI-30827 attenuated Tat-induced inhibition of [3H]WIN35,428 binding. These observations demonstrate that tyrosine 470 and 88 are critical for allosteric modulatory effects of SRI-compounds on the interaction of cocaine with hDAT.