Mutations of Human DopamineTransporter at Tyrosine88, Aspartic Acid206, and Histidine547 Influence Basal and HIV-1 Tat-inhibited Dopamine Transport.
Mutations of Human DopamineTransporter at Tyrosine88, Aspartic Acid206, and Histidine547 Influence Basal and HIV-1 Tat-inhibited Dopamine Transport.
复制标题
DOI：
10.1007/s11481-021-09984-5
复制
发表时间：
2021-12
期刊：
影响因子：
--
通讯作者：
Zhu J
中科院分区：
文献类型：
--
作者：
Quizon PM;Yuan Y;Zhu Y;Zhou Y;Strauss MJ;Sun WL;Zhan CG;Zhu J
HIV-1转录反式激活因子（达特）通过阻断多巴胺的传递，对HIV-1相关性神经认知障碍的发生发展有重要影响。本研究确定了人多巴胺转运蛋白（hDAT）对基础和Tat诱导的多巴胺转运抑制的突变效应。与野生型hDAT相比，D381 L和Y88 F/D206 L/H547 A中[3 H]多巴胺摄取的最大速度（Vmax）降低，D206 L/H547 A中增加，而D206 L中无变化。重组TatR 1 − 86抑制野生型hDAT中多巴胺的摄取，这在DAT突变体（D206 L，D206 L/H547 A和Y88 F/D206 L/H547 A）或突变的TatR 1 − 86（K19 A和C22 G）中减弱，表明Tat-DAT相互作用受到干扰。hDAT对转运蛋白构象的突变效应通过减弱D206 L/H547 A和Y88 F/D206 A/H547 A中锌诱导的增加的[3 H] WIN 35，428结合以及增强D206 L/H547 A中的基础MPP+流出来证明。H547 A诱导的向外开放转运构象状态通过在hDAT背景上增强的对插入的半胱氨酸（I159 C）的MTSET（[2-（三甲基铵）乙基]-甲硫基磺酸盐）的可及性来进一步验证。此外，H547 A显示棕榈酰化修饰物诱导的多巴胺摄取抑制相对于野生型hDAT增加，表明H547 A中基础棕榈酰化的变化。这些结果表明，Y88 F，D206 L，和H547 A减弱达特抑制，同时保持DA摄取，提供了深入了解，以确定改善DAT介导的多巴胺能失调的目标。HIV-1达特通过直接变构相互作用抑制突触前末梢上的人多巴胺转运蛋白（hDAT）的多巴胺摄取。预测关键hDAT残基D-H547、D-Y88和D-D206参与HIV-1 Tat-DAT结合。突变这些残基通过破坏Tat-hDAT相互作用减弱这种抑制作用。在线版本包含补充材料，可在10.1007/s11481-021-09984-5获得。
HIV-1 transactivator of transcription (Tat) has a great impact on the development of HIV-1 associated neurocognitive disorders through disrupting dopamine transmission. This study determined the mutational effects of human dopamine transporter (hDAT) on basal and Tat-induced inhibition of dopamine transport. Compared to wild-type hDAT, the maximal velocity (Vmax) of [3H]dopamine uptake was decreased in D381L and Y88F/D206L/H547A, increased in D206L/H547A, and unaltered in D206L. Recombinant TatR1 − 86 inhibited dopamine uptake in wild-type hDAT, which was attenuated in either DAT mutants (D206L, D206L/H547A, and Y88F/D206L/H547A) or mutated TatR1 − 86 (K19A and C22G), demonstrating perturbed Tat-DAT interaction. Mutational effects of hDAT on the transporter conformation were evidenced by attenuation of zinc-induced increased [3H]WIN35,428 binding in D206L/H547A and Y88F/D206A/H547A and enhanced basal MPP+ efflux in D206L/H547A. H547A-induced outward-open transport conformational state was further validated by enhanced accessibility to MTSET ([2-(trimethylammonium)ethyl]-methanethiosulfonate) of an inserted cysteine (I159C) on a hDAT background.. Furthermore, H547A displayed an increase in palmitoylation inhibitor-induced inhibition of dopamine uptake relative to wide-type hDAT, indicating a change in basal palmitoylation in H547A. These results demonstrate that Y88F, D206L, and H547A attenuate Tat inhibition while preserving DA uptake, providing insights into identifying targets for improving DAT-mediated dopaminergic dysregulation. HIV-1 Tat inhibits dopamine uptake through human dopamine transporter (hDAT) on the presynaptic terminal through a direct allosteric interaction. Key hDAT residues D-H547, D-Y88, and D-D206 are predicted to be involved in the HIV-1 Tat-DAT binding. Mutating these residues attenuates this inhibitory effect by disrupting the Tat-hDAT interaction The online version contains supplementary material available at 10.1007/s11481-021-09984-5.
登录
查看更多内容
影响因子：
5.7
作者：
Chang, Linda;Wang, Gene-Jack;Fowler, Joanna S.
通讯作者：
Fowler, Joanna S.
影响因子：
3.2
作者：
Heaton, Robert K.;Franklin, Donald R.;Ellis, Ronald J.;McCutchan, J. Allen;Letendre, Scott L.;LeBlanc, Shannon;Corkran, Stephanie H.;Duarte, Nichole A.;Clifford, David B.;Woods, Steven P.;Collier, Ann C.;Marra, Christina M.;Morgello, Susan;Mindt, Monica Rivera;Taylor, Michael J.;Marcotte, Thomas D.;Atkinson, J. Hampton;Wolfson, Tanya;Gelman, Benjamin B.;McArthur, Justin C.;Simpson, David M.;Abramson, Ian;Gamst, Anthony;Fennema-Notestine, Christine;Jernigan, Terry L.;Wong, Joseph;Grant, Igor
通讯作者：
Grant, Igor
影响因子：
6.1
作者：
Gaskill PJ;Miller DR;Gamble-George J;Yano H;Khoshbouei H
通讯作者：
Khoshbouei H
影响因子：
5
作者：
Loland, CJ;Norgaard-Nielsen, K;Gether, U
通讯作者：
Gether, U
影响因子：
6.2
作者：
Midde, Narasimha M.;Huang, Xiaoqin;Zhu, Jun
通讯作者：
Zhu, Jun