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: 8073152
• 负责人: JEFFRY D. MADURA
• 金额: $ 29.68万
• 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/8073152
• 关键词: 3-Dimensional; Antidepressive Agents; Anxiety Disorders; Applications Grants; Area; Argentina; 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; Mental Depression; Methylphenidate; Modeling; Molecular; Molecular Conformation; Molecular Models; Mutagenesis; Narcolepsy; Nervous system structure; Neurosciences; Parkinson Disease; Process; Proteins; Psychostimulant dependence; Publishing; Quantitative Structure-Activity Relationship; Research; Research Activity; Research Infrastructure; Scientist; Serotonin; Structure; Students; Substance Addiction; Substance abuse problem; Techniques; Testing; Textbooks; Therapeutic; Training; Transmembrane Domain; United States; Work; base; chronic pain; combat; comparative; computer studies; design; dopamine transporter; 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）：这项CRCNS拨款申请提出了质膜多巴胺（DAT）和血清素转运体（SERT）的结构、功能和动态研究。最近，随着同源LeuTAa亮氨酸转运体的结晶，鉴定DAT和SERT底物和抑制剂结合位点的艰巨过程得到了意想不到的推动。该晶体结构揭示了与亮氨酸底物相邻的跨膜（TMs） 1和6的铰接区域，TMs 3、8和10也描绘了结合袋。通过比较分子建模技术，我们利用LeuTAa发表了DAT的三维模型。该模型还提出了新的抑制剂结合位点，与多巴胺不同，可以通过分子药理学技术进行测试。智力优势：该项目汇集了计算科学家、药物化学家和药理学家组成的独特团队，研究单胺类神经转运蛋白（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兴奋剂和抗抑郁药抑制剂的结合位置，以及参与转运机制的构象状态。对接、先进分子动力学模拟、QSAR和基于结构的设计领域的最先进的计算技术将用于识别转运蛋白的重要残基和区域，以进行诱变实验，以及指导合成抑制非神经递质分子结合但保持转运蛋白活性的新化合物。更广泛的影响：该提案的影响分为三类：(1)科学界，(2)机构，(3)个人研究小组。通过验证和测试DAT和SERT的三维结构的可用性，科学界将受益于该项目的结果。这些结构可以用于基于结构的设计，这是目前基于配体的设计工作的一个进步。由此产生的3-D转运体“蓝图”可用于筛选新的MAT配体数据库。可以阐明特定的MAT蛋白质成分如何有助于区分可卡因（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，因为寻找治疗上有用的化合物来对抗精神兴奋剂滥用和成瘾已经进行了几十年，迄今为止还没有临床可用的药物。这项工作可能为合理设计mat相关疾病的治疗方法打开大门，这些疾病包括药物滥用和成瘾、抑郁症、焦虑症、注意力缺陷多动障碍、嗜睡症、慢性疼痛和帕金森病。该社区还将受益于在计算和实验神经科学方面交叉训练的新毕业科学家的加入。对该机构的影响将是增加研究基础设施和促进多学科研究活动。这项建议对个别研究小组的影响将是加强他们从有才能和多样化的学生中选择的努力。目前，合并后的小组有5名男生和4名女生。这些学生来自阿根廷、中国、印度和美国。我们还将继续提供分子建模课程，该研究小组的几名现任学生已经上过这门课程。我们的经验和成果将为分子建模课程提供新的材料。最后，PI是普通化学教科书的作者，这项工作的结果将被纳入教科书。综上所述，该提案符合以下标准：(1)它包括计算和/或建模专家（Madura研究组），实验神经药理学家（Surratt研究组）和药物化学家（Lapinsky研究组）之间的合作；(2)在多巴胺和血清素神经转运体的模型开发和完善方面，这些研究小组之间的合作涉及激烈的、动态的互动；(3)引导新模型的开发和测试，为实验设计和新假设的产生提供框架，以揭示神经系统正常或患病状态的潜在机制。