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: 7869264
• 负责人: JEFFRY D. MADURA
• 金额: $ 30.6万
• 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/7869264
• 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; Mental Depression; 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; 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）：该 CRCNS 拨款申请提出了对质膜多巴胺 (DAT) 和血清素转运蛋白 (SERT) 的结构、功能和动态研究。最近，随着同源 LeuTAa 亮氨酸转运蛋白的结晶，鉴定 DAT 和 SERT 底物及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 (TM) 1 和 6 的铰接区域，TM 3、8 和 10 也描绘了结合袋。通过比较分子建模技术，我们发布了使用 LeuTAa 的 DAT 三维模型。该模型还提出了与多巴胺不同的新型抑制剂结合位点，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的独特团队，研究单胺神经转运蛋白（MAT）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抑制剂的结合位置，以及转运机制中涉及的构象状态。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直接合成抑制非神经递质分子结合但保留转运蛋白活性的新型化合物。更广泛的影响：该提案的影响分为三类：(1) 科学界，(2) 机构，(3) 个别研究小组。通过提供经过验证和测试的 DAT 和 SERT 三维结构，科学界将从该项目的成果中受益。然后，这些结构可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。由此产生的 3-D 转运蛋白“蓝图”可用于筛选新型 MAT 配体的数据库。可以阐明特定的 MAT 蛋白成分如何有助于区分可卡因（高滥用可能性）和哌醋甲酯（低滥用可能性）。这是一个至关重要的健康问题，因为数十年来一直在寻找治疗上有用的化合物来对抗精神兴奋剂滥用和成瘾，但迄今为止还没有临床可用的药物。拟议的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、发作性睡病、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领域接受交叉培训的新毕业科学家的加入。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。该提案对各个研究团队的影响将是加强他们从才华横溢且多元化的学生群体中进行选拔的努力。目前，合并后的小组有五名男学生和四名女学生。这些学生来自阿根廷、中国、印度和美国。我们还将继续提供分子建模课程，研究团队的一些现有学生已经修读了该课程。分子建模课程的新材料将取自我们在该项目中的经验和成果。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。总之，该提案在以下方面符合该计划的标准：（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) 这项合作涉及这些研究小组在多巴胺和血清素神经转运蛋白的模型开发和完善方面进行的激烈、动态的互动； （3）导致新模型的开发和测试，为实验设计和新假设的生成提供框架，以揭示神经系统正常或患病状态下的机制。