MT COBRE: MAPPING SEROTONIN TRANSPORTER BINDING DOMAINS

MT COBRE：绘制血清素转运蛋白结合域图

• 批准号: 7720402
• 负责人: JOHN M GERDES
• 金额: $ 14.59万
• 依托单位: UNIVERSITY OF MONTANA
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-05-01 至 2009-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7720402
• 关键词: Amino Acids; Binding; Binding Sites; Carrier Proteins; Chemicals; Collection; Computer Retrieval of Information on Scientific Projects Database; Computer Simulation; Custom; Fire - disasters; Funding; Goals; Grant; Head; Homo; Institution; Integral Membrane Protein; Label; Ligand Binding; Ligands; Link; Maps; Mass Spectrum Analysis; Measurement; Mental disorders; Modeling; Mono-S; Neurologic; Neurons; Neurotransmitters; Operative Surgical Procedures; Protein Binding; Quipazine; Research; Research Personnel; Resources; Sequence Analysis; Serotonin; Site; Source; Structure; Synaptic Cleft; Tertiary Protein Structure; Thinking; United States National Institutes of Health; base; crosslink; design; receptor density; serotonin transporter; three dimensional structure

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The serotonin transporter (SERT) is an integral membrane protein responsible for clearance of the endogenous biogenic neurotransmitter 5-HT from the synaptic cleft after neuronal firing. Abnormalities in SERT structure-function and/or changes in SERT receptor densities have been implicated in a variety of significant neurological and mental health disorders. Thus, an enhanced understanding of SERT operation is needed. The exact details of SERT global tertiary structure and intimate three-dimensional (3D) representations of critical protein binding regions linked to the SERT transport phenomena remain ill defined. The SERT structure organization is thought to possess a ligand binding site that is composed by amino acids assembled in 3D space by the SERT tertiary protein structure. It is our hypothesis that the efficacious mapping of the amino acid constituents of the SERT binding domains will reveal critical features of the SERT tertiary structure. Our long-term objective is to significantly contribute to understanding of the commonalities and differences amongst key CNS transport proteins in terms of structure-function. The original goal of this project has been to map participating amino acid residues within critical SERT binding domains with custom designed, potent and selective, quipazine-based cross-linking probes. The following three specific aims will be accomplished: 1. To construct a 3D computational model of SERT that will enable accurate measurements of inter-residue secondary structure. 2. To synthesize and pharmacologically characterize a collection of mono- and homo-bifunctional probes defined by our computationally derived ligand design criteria. 3. To determine the amino acid cross-linked SERT 3D structure maps, first by the irreversible chemical tagging of the dual-headed cross-linking agents to expressed SERT, followed by mass spectrometry tagged SERT sequence analysis, and then correlating the labeled residue sites with the 3D SERT model.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 5-羟色胺转运蛋白（SERT）是一种整体膜蛋白，负责从神经元射击后突触裂口清除内源性生物源性神经递质5-HT。 SERT结构功能的异常和/或SERT受体密度的变化已与各种重要的神经系统疾病有关。因此，需要增强对SERT操作的理解。 SERT全球三级结构的确切细节和与SERT转运现象相关的关键蛋白结合区域的亲密三维表示。 SERT结构组织被认为具有由Sert三级蛋白质结构在3D空间组装的氨基酸组成的配体结合位点。我们的假设是，SERT结合结构域的氨基酸成分的有效映射将揭示Sert三级结构的关键特征。我们的长期目标是在结构功能方面显着有助于理解关键中枢神经系统运输蛋白的共同性和差异。 该项目的最初目标是用定制，有效的，基于quipazine的交联探针在关键SERT结合域中绘制参与参与的氨基酸残基。以下三个特定目标将实现： 1。构建SERT的3D计算模型，该模型将实现精确测量二人间二级结构。 2。在合成和药理学上表征由我们的计算衍生的配体设计标准定义的单官能和同源函数探针的集合。 3。为了确定氨基酸交联SERT 3D结构图，首先是由双头交联的不可逆化学标记与表达的SERT的不可逆化学标记，然后进行质谱标记的SERT序列分析，然后将标记的残基位点与3D Sert模型相关联。