CRCNS: Computational and experimental study of dopamine and serotonin transporter

CRCNS：多巴胺和血清素转运蛋白的计算和实验研究

• 批准号: 7771845
• 负责人: JEFFRY D. MADURA
• 金额: $ 30.91万
• 依托单位: DUQUESNE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-01 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7771845
• 关键词: 3-Dimensional; Antidepressive Agents; Anxiety Disorders; Applications Grants; Area; Argentina; Arts; Attention deficit hyperactivity disorder; Binding; Binding Sites; Categories; Cell membrane; Chemistry; China; Cocaine; Collaborations; Communities; Computational Technique; Crystallization; Databases; Development; Discrimination; Docking; Dopamine; Female; Generations; Goals; Health; India; Individual; Institution; Leucine; Ligands; Location; Madura; Methylphenidate; Modeling; Molecular; Molecular Conformation; Molecular Models; Mutagenesis; Narcolepsy; Nervous system structure; Neurosciences; Neurotransmitters; Parkinson Disease; Process; Proteins; Publishing; Quantitative Structure-Activity Relationship; Research; Research Activity; Research Infrastructure; Scientist; Serotonin; Structure; Students; Substance abuse problem; Techniques; Testing; Textbooks; Therapeutic; Training; Transmembrane Domain; United States; Work; addiction; base; chronic pain; combat; comparative; depression; design; experience; falls; inhibitor/antagonist; male; model development; molecular dynamics; molecular modeling; monoamine; neurotransmitter transport; novel; programs; psychostimulant; research study; serotonin transporter; stimulant abuse; three dimensional structure; three-dimensional modeling

DESCRIPTION (provided by applicant): This CRCNS grant application proposes structure, function, and dynamic studies on the plasma membrane dopamine (DAT) and serotonin transporters (SERT). Recently, the arduous process of identifying DAT and SERT substrate and inhibitor binding sites received an unexpected boost with the crystallization of the homologous LeuTAa leucine transporter. This crystal structure revealed hinged regions of transmembranes (TMs) 1 and 6 adjacent to the leucine substrate, with TMs 3, 8 and 10 also delineating the binding pocket. Through comparative molecular modeling techniques we have published a three-dimensional model of DAT using LeuTAa. The modeling also suggests novel inhibitor binding sites, nonidentical to dopamine, that can be tested via molecular pharmacological techniques. Intellectual Merit: This project brings together a unique team of computational scientists, medicinal chemists, and pharmacologists to examine the structure, function, and dynamics of monoamine neurotransporters (MATs). The overall goal of this project is to determine binding locations for psychostimulant and antidepressant inhibitors of neurotransmitter transport, and the conformational states involved in the transport mechanism. State-of-the-art computational techniques in areas of docking, advanced molecular dynamics simulations, QSAR and structure-based design will be used to identify important residues and regions of the transporter to perform mutagenesis experiments as well as the direct the synthesis of novel compounds that inhibit binding of non-neurotransmitter molecules yet retain transporter activity. Broader Impacts: The impacts of this proposal fall into three categories: (1) the scientific community, (2) the institution, and (3) the individual research groups. The scientific community will benefit from the results of this project through the availability of validated and tested three-dimensional structures of DAT and SERT. These structures can then be used in structure-based design, which is an advance on current ligand-based design efforts. The resultant 3-D transporter "blueprint" can be used to screen databases for novel MAT ligands. How specific MAT protein components contribute to discrimination between, for example, cocaine (high abuse potential) and methylphenidate (low abuse potential) may be elucidated. This is a critical health issue, as the search for therapeutically useful compounds in combating psychostimulant abuse and addiction has been in progress for decades, with no clinically available agents to date. The proposed work may open the door to rational design of therapeutics for MAT-related conditions that include substance abuse and addiction, depression, anxiety disorders, attention deficit hyperactivity disorder, narcolepsy, chronic pain and Parkinson's disease. The community will also benefit from the addition of newly graduated scientists cross-trained in computational and experimental neurosciences. The impact at the institution will be increased research infrastructure and promotion of multi-disciplinary research activities. The impact of this proposal on the individual research teams will be to enhance their efforts to select from a talented and diverse pool of students. Currently the combined groups have five male and four female students. These students come from Argentina, China, India, and the United States. We will also continue to offer a molecular modeling course in which several of the current students of the research team have already taken. New material for the molecular modeling course will be drawn from our experiences and results from this project. Finally the PI is a general chemistry textbook author and results from this work will be incorporated into the textbook. In summary, this proposal fits the criteria for this program in the following manner: (1) it includes collaborations between computational and/or modeling experts (Madura research group), experimental neuropharmacologists (Surratt research group) and medicinal chemists (Lapinsky research group); (2) this collaboration involves intense, dynamic interactions among these research groups in the model development and refinement of dopamine and serotonin neurotransporters; and (3) leading to the development and testing of new models that provide a framework for the design of experiments and the generation of new hypotheses to reveal mechanisms underlying normal or diseased states of the nervous system.

描述（由申请人提供）：此CRCN授予应用程序提出了有关质膜多巴胺（DAT）和5-羟色胺转运蛋白（SERT）的结构，功能和动态研究。最近，鉴定DAT和SERT底物和抑制剂结合位点的艰巨过程通过同源Leutaa Leucine转运蛋白的结晶而产生了意外的提升。这种晶体结构揭示了与亮氨酸基板相邻的跨膜（TMS）1和6的铰链区域，TMS 3、8和10也描绘了结合口袋。通过比较分子建模技术，我们发布了使用Leutaa的DAT的三维模型。该建模还表明，可以通过分子药理技术对多巴胺非相同的新型抑制剂结合位点进行测试。知识分子的优点：该项目汇集了一个独特的计算科学家，药物学家和药理学家的团队，以检查单胺神经转运蛋白（MATS）的结构，功能和动力学。该项目的总体目标是确定神经递质转运的精神刺激剂和抗抑郁剂抑制剂的结合位置，以及涉及运输机制的构象状态。最先进的计算技术在对接，高级分子动力学模拟，QSAR和基于结构的设计领域的最新计算技术将用于识别运输蛋白的重要残基和区域，以执行诱变实验，以及直接合成新型化合物的新型化合物，这些化合物抑制非凝聚递质分子的结合，并导致无效分子的结合。更广泛的影响：该提案的影响分为三类：（1）科学界，（2）机构和（3）个别研究小组。科学界将通过验证和测试的DAT和SERT的三维结构的可用性从该项目的结果中受益。然后可以将这些结构用于基于结构的设计，这是当前基于配体的设计工作的进步。所得的3-D转运蛋白“蓝图”可用于筛选新型MAT配体的数据库。特定的MAT蛋白成分如何有助于歧视可卡因（高滥用潜力）和哌醋甲酯（低滥用潜力）。这是一个关键的健康问题，因为几十年来一直在进行对抗精神刺激和成瘾的治疗效果，迄今为止尚无临床可用的代理。拟议的工作可能为与MAT相关的疾病的理性设计打开了大门，包括药物滥用和成瘾，抑郁症，焦虑症，注意力缺陷多动障碍，发毒症，发毒症，慢性疼痛和帕金森氏病。在计算和实验性神经科学中，新毕业的科学家的添加还将受益。该机构的影响将增加研究基础设施和促进多学科研究活动。该提案对单个研究团队的影响将是加强他们从有才华和多样化的学生中选择的努力。目前，合并后的小组有五名男性和四名女学生。这些学生来自阿根廷，中国，印度和美国。我们还将继续提供一门分子建模课程，其中现有研究团队的几个学生已经参加了该课程。分子建模课程的新材料将从我们的经验和该项目的结果中获取。最后，PI是一家通用化学教科书作者，这项工作的结果将被纳入教科书中。总而言之，该提案以以下方式符合该计划的标准：（1）它包括计算和/或建模专家（Madura Research Group），实验性神经药物学家（Surratt Research Group）和药物化学家（Lapinsky Research Group）之间的合作； （2）这种合作涉及这些研究小组之间在多巴胺和5-羟色胺神经递送蛋白的改进中进行激烈的动态相互作用； （3）导致开发和测试新模型，这些新模型为实验设计和生成新的假设提供了一个框架，以揭示神经系统正常状态或患病状态的机制。