A Novel Biotinylated Homotryptamine Derivative for Quantum Dot Imaging of Serotonin Transporter in Live Cells.
A Novel Biotinylated Homotryptamine Derivative for Quantum Dot Imaging of Serotonin Transporter in Live Cells.
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DOI：
10.3389/fncel.2021.667044
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发表时间：
2021
影响因子：
5.3
通讯作者：
Rosenthal SJ
中科院分区：
文献类型：
--
作者：
Tomlinson ID;Kovtun O;Torres R;Bellocchio LG;Josephs T;Rosenthal SJ
5-羟色胺转运体（SERT）是选择性5-羟色胺再摄取抑制剂（SSRI）抗抑郁药的主要靶点，这些抗抑郁药被认为通过增加5-羟色胺的突触浓度来发挥其治疗作用。因此，可用于研究SERT的细胞运输的探针是有价值的研究工具。我们已经开发了一种新的配体（IDT 785），它是由一个SERT拮抗剂（四氢吡啶吲哚衍生物）共轭的生物素化的聚乙二醇（PEG）通过苯乙基接头。该化合物具有生物活性，可抑制SERT介导的IDT 307再摄取，半最大抑制浓度为7.2 ± 0.3 μM。我们证明了IDT 785能够在转染的HEK-293培养物中对膜SERT进行量子点（QD）标记，这可以使用高亲和力5-羟色胺再摄取抑制剂帕罗西汀进行阻断。分子对接研究表明，IDT 785可能是结合到细胞外前庭结合位点，而不是orthosteric底物结合位点，这可能是由于PEG链的亲水性和增加的自由度的损失，将需要渗透到orthosteric结合位点。使用IDT 785，我们能够研究HEK-293细胞中异源表达的YFP-SERT的膜定位和膜动力学，并证明SERT表达富集在膜边缘和薄的细胞突起中。
The serotonin transporter (SERT) is the primary target for selective serotonin reuptake inhibitor (SSRI) antidepressants that are thought to exert their therapeutic effects by increasing the synaptic concentration of serotonin. Consequently, probes that can be utilized to study cellular trafficking of SERT are valuable research tools. We have developed a novel ligand (IDT785) that is composed of a SERT antagonist (a tetrahydro pyridyl indole derivative) conjugated to a biotinylated poly ethylene glycol (PEG) via a phenethyl linker. This compound was determined to be biologically active and inhibited SERT-mediated reuptake of IDT307 with the half-maximal inhibitory concentration of 7.2 ± 0.3 μM. We demonstrated that IDT785 enabled quantum dot (QD) labeling of membrane SERT in transfected HEK-293 cultures that could be blocked using the high affinity serotonin reuptake inhibitor paroxetine. Molecular docking studies suggested that IDT785 might be binding to the extracellular vestibule binding site rather than the orthosteric substrate binding site, which could be attributable to the hydrophilicity of the PEG chain and the increased loss of degrees of freedom that would be required to penetrate into the orthosteric binding site. Using IDT785, we were able to study the membrane localization and membrane dynamics of YFP-SERT heterologously expressed in HEK-293 cells and demonstrated that SERT expression was enriched in the membrane edge and in thin cellular protrusions.
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影响因子：
--
作者：
COOL, DR;LEIBACH, FH;GANAPATHY, V
通讯作者：
GANAPATHY, V
影响因子：
64.8
作者：
Coleman, Jonathan A.;Yang, Dongxue;Gouaux, Eric
通讯作者：
Gouaux, Eric
DOI：
10.1523/jneurosci.0048-12.2012
发表时间：
2012-06-27
期刊：
The Journal of neuroscience : the official journal of the Society for Neuroscience
影响因子：
--
作者：
Chang JC;Tomlinson ID;Warnement MR;Ustione A;Carneiro AM;Piston DW;Blakely RD;Rosenthal SJ
通讯作者：
Rosenthal SJ
影响因子：
64.8
作者：
BLAKELY, RD;BERSON, HE;BRADLEY, CC
通讯作者：
BRADLEY, CC
差异5-羟色胺转运与外周血细胞中的RH5-HTTLPR有关。
影响因子：
6.8
作者：
Singh YS;Altieri SC;Gilman TL;Michael HM;Tomlinson ID;Rosenthal SJ;Swain GM;Murphey-Corb MA;Ferrell RE;Andrews AM
通讯作者：
Andrews AM