Structural Dynamics of the Human Serotonin Transporter
人类血清素转运蛋白的结构动力学
基本信息
- 批准号:10386296
- 负责人:
- 金额:$ 3.21万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2021
- 资助国家:美国
- 起止时间:2021-12-01 至 2024-11-30
- 项目状态:已结题
- 来源:
- 关键词:3-DimensionalAmericanAmphetamine AddictionAmphetaminesBenchmarkingBiochemicalBrainCell membraneCellsChloride IonCollaborationsCoupledCryoelectron MicroscopyCrystallizationCrystallographyDataData AnalysesDimensionsDiseaseDopamineDrosophila melanogasterDrug DesignElectron MicroscopyElectron Spin Resonance SpectroscopyElectronsElementsEngineeringEquilibriumFingerprintFoundationsFunctional disorderFutureGoalsHumanIon CotransportKnowledgeLabelLaboratoriesLengthLigandsLightLinkLipidsMediatingMental DepressionMental disordersMentorshipMethodologyModelingMolecularMolecular ConformationMutationN-terminalNeuronsNeurotransmittersNorepinephrinePhosphatidylinositol 4,5-DiphosphatePhysiologicalProteinsResearchResolutionRoleSamplingSerotoninSignal TransductionSiteSodiumSolventsSpectrum AnalysisSpin LabelsStimulantStructureSurveysSynapsesTestingTransmembrane Domainabuse liabilityaddictionautism spectrum disordercomputer studiesconformational conversioncryogenicsdata acquisitiondopamine transporterdrug actiondrug of abuseecstasyexperimental studyin vivoinsightmemberneuropsychiatric disorderneurotransmitter transportnovel therapeuticspresynapticprotein expressionprotein protein interactionprotein purificationreuptakeserotonin transporterskillssodium ionsuccesssymportertargeted treatmenttherapeutic developmenttool
项目摘要
PROJECT SUMMARY
As a member of the class of neurotransmitter:sodium symporters (NSSs), the serotonin transporter (SERT)
regulates levels of serotonin in the brain through reuptake or forward transport. Dysfunction of SERT has been
associated with neuropsychiatric disorders and autism in humans. As such, SERT is a major target for
therapeutic development as well as sites of action of drugs of abuse. Transport of serotonin from the synapse
into the presynaptic cell is energetically coupled to symport of sodium and chloride ions. However, in the
presence of amphetamine derivatives, this mechanism is altered. Molecules in the amphetamine class induce
reverse transport by NSSs, leading to increased synaptic levels of serotonin, dopamine, and norepinephrine.
This reverse transport mechanism is mediated by the N-terminus of the transporter, a hub for protein-protein
interactions and signaling, and is modulated by phosphatidylinositol-4,5-bisphosphate (PIP2), a lipid enriched in
the plasma membrane of neurons. The central objective of this proposal is to examine the conformational
dynamics underlying transport for human SERT (hSERT), illuminating the role of its N-terminus in both
forward and reverse transport. Aim 1 will use double electron-electron resonance (DEER) spectroscopy to
probe conformational equilibria of full-length hSERT under different conditions to benchmark structural
transitions across the transport cycle. Aim 2 will characterize the conformation of the N-terminus through
surveying the mobility and solvent accessibility of singly labeled sites and define its structural reorganization in
the presence of PIP2. The N-terminus and PIP2 are hypothesized to impose structural changes to the hSERT
transmembrane domain, which prompts reverse transport in the presence of amphetamine. The proposed
experiments are expected to reveal insights into the transport mechanism of eukaryotic NSSs and could be
foundational for understanding how the presence of disease-linked mutations or exogenous molecules such as
amphetamine could disturb this mechanism.
The laboratory of Dr. Hassane Mchaourab specializes in revealing the conformational dynamics of
transporters, including bacterial homologs of NSS, utilizing the tools of electron paramagnetic resonance (EPR)
Our collaborator, Dr. Eric Gouaux has expertise in the field of eukaryotic NSS proteins and has determined
several high-resolution structures of hSERT. Together, the mentorship of Dr. Mchaourab and the guidance of
Dr. Gouaux will enable the success of experiments outlined in my aims from mammalian protein expression and
purification through EPR data acquisition and analysis and culminating in mechanistic models that integrate high-
resolution structures with conformational dynamics.
项目总结
作为神经递质的一员:钠转运体(NSSS),5-羟色胺转运体(SERT)
通过重新摄取或前向转运来调节大脑中的5-羟色胺水平。SERT功能障碍一直以来
与人类的神经精神障碍和自闭症有关。因此,SERT是
滥用药物的治疗发展和作用部位。5-羟色胺从突触的运输
进入突触前细胞与钠离子和氯离子的结合是能量耦合的。然而,在
在苯丙胺衍生物的存在下,这一机制被改变。安非他明类分子能诱导
NSSS的反向转运,导致5-羟色胺、多巴胺和去甲肾上腺素的突触水平增加。
这种反向转运机制是由转运蛋白的N-末端介导的,转运蛋白是蛋白质-蛋白质的枢纽
相互作用和信号传递,并受磷脂酰肌醇-4,5-二磷酸(PIP2)调节,PIP2是一种富含
神经元的质膜。这项建议的中心目标是研究构象
人类SERT(HSERT)转运的动力学基础,阐明其N末端在两者中的作用
正向和反向传输。目标1将使用双电子-电子共振(DeER)光谱来
探索全长hSERT在不同条件下的构象平衡以作为结构基准
在整个运输周期中的过渡。目标2将通过以下方式表征N末端的构象
调查单一标记位点的流动性和溶剂可及性,并确定其结构重组
PIP2的存在。N末端和PIP2被假设为对hSERT施加结构变化
跨膜结构域,在苯丙胺存在时促使反向转运。建议数
实验有望揭示真核细胞NSSS的运输机制,并可能
对于理解与疾病相关的突变或外源分子(如
安非他命可能会扰乱这种机制。
Hassane Mchaourab博士的实验室专门研究
转运体,包括NSS的细菌同源物,利用电子顺磁共振(EPR)工具
我们的合作者Eric Gouaux博士在真核细胞NSS蛋白领域拥有专业知识,并已确定
HSERT的几种高分辨率结构。总而言之,Mchaourab博士的指导和
Gouaux博士将使我的目标中概述的实验从哺乳动物蛋白质表达和
通过EPR数据采集和分析进行提纯,并最终形成集成了高
具有构象动力学的分辨结构。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
数据更新时间:{{ journalArticles.updateTime }}
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
数据更新时间:{{ journalArticles.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ monograph.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ sciAawards.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ conferencePapers.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ patent.updateTime }}
Alexandra Corinne Schwartz其他文献
Alexandra Corinne Schwartz的其他文献
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
{{ truncateString('Alexandra Corinne Schwartz', 18)}}的其他基金
Structural Dynamics of the Human Serotonin Transporter
人类血清素转运蛋白的结构动力学
- 批准号:
10610326 - 财政年份:2021
- 资助金额:
$ 3.21万 - 项目类别:
相似海外基金
Collaborative Research: REU Site: Earth and Planetary Science and Astrophysics REU at the American Museum of Natural History in Collaboration with the City University of New York
合作研究:REU 地点:地球与行星科学和天体物理学 REU 与纽约市立大学合作,位于美国自然历史博物馆
- 批准号:
2348998 - 财政年份:2025
- 资助金额:
$ 3.21万 - 项目类别:
Standard Grant
Collaborative Research: REU Site: Earth and Planetary Science and Astrophysics REU at the American Museum of Natural History in Collaboration with the City University of New York
合作研究:REU 地点:地球与行星科学和天体物理学 REU 与纽约市立大学合作,位于美国自然历史博物馆
- 批准号:
2348999 - 财政年份:2025
- 资助金额:
$ 3.21万 - 项目类别:
Standard Grant
Collaborative Research: Ionospheric Density Response to American Solar Eclipses Using Coordinated Radio Observations with Modeling Support
合作研究:利用协调射电观测和建模支持对美国日食的电离层密度响应
- 批准号:
2412294 - 财政年份:2024
- 资助金额:
$ 3.21万 - 项目类别:
Standard Grant
Conference: Doctoral Consortium at Student Research Workshop at the Annual Conference of the North American Chapter of the Association for Computational Linguistics (NAACL)
会议:计算语言学协会 (NAACL) 北美分会年会学生研究研讨会上的博士联盟
- 批准号:
2415059 - 财政年份:2024
- 资助金额:
$ 3.21万 - 项目类别:
Standard Grant
Conference: Polymeric Materials: Science and Engineering Division Centennial Celebration at the Spring 2024 American Chemical Society Meeting
会议:高分子材料:美国化学会 2024 年春季会议科学与工程部百年庆典
- 批准号:
2415569 - 财政年份:2024
- 资助金额:
$ 3.21万 - 项目类别:
Standard Grant
Collaborative Research: RUI: Continental-Scale Study of Jura-Cretaceous Basins and Melanges along the Backbone of the North American Cordillera-A Test of Mesozoic Subduction Models
合作研究:RUI:北美科迪勒拉山脊沿线汝拉-白垩纪盆地和混杂岩的大陆尺度研究——中生代俯冲模型的检验
- 批准号:
2346565 - 财政年份:2024
- 资助金额:
$ 3.21万 - 项目类别:
Standard Grant
REU Site: Research Experiences for American Leadership of Industry with Zero Emissions by 2050 (REALIZE-2050)
REU 网站:2050 年美国零排放工业领先地位的研究经验 (REALIZE-2050)
- 批准号:
2349580 - 财政年份:2024
- 资助金额:
$ 3.21万 - 项目类别:
Standard Grant
Collaborative Research: RUI: Continental-Scale Study of Jura-Cretaceous Basins and Melanges along the Backbone of the North American Cordillera-A Test of Mesozoic Subduction Models
合作研究:RUI:北美科迪勒拉山脊沿线汝拉-白垩纪盆地和混杂岩的大陆尺度研究——中生代俯冲模型的检验
- 批准号:
2346564 - 财政年份:2024
- 资助金额:
$ 3.21万 - 项目类别:
Standard Grant
Conference: Latin American School of Algebraic Geometry
会议:拉丁美洲代数几何学院
- 批准号:
2401164 - 财政年份:2024
- 资助金额:
$ 3.21万 - 项目类别:
Standard Grant
Conference: North American High Order Methods Con (NAHOMCon)
会议:北美高阶方法大会 (NAHOMCon)
- 批准号:
2333724 - 财政年份:2024
- 资助金额:
$ 3.21万 - 项目类别:
Standard Grant