Dynamics of the Opioid Receptor Signaling Complex

阿片受体信号复合物的动力学

• 批准号: 8944883
• 负责人: Kurt Wuthrich
• 金额: $ 37.43万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-01 至 2019-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8944883
• 关键词: Acute Pain; Address; Adrenergic Receptor; Adverse effects; Affinity; Agonist; Alcohols; Amphetamines; Behavior; Behavioral; Binding; Biochemistry; Biological Assay; California; Cessation of life; Cocaine; Complex; Coupled; Coupling; Data; Development; Drug Formulations; Drug Prescriptions; Drug Targeting; Dynorphins; Engineering; Evaluation; Family; France; Funding; Funding Applicant; Future; G-Protein-Coupled Receptors; GTP-Binding Proteins; Gatekeeping; Goals; Growth; Human; Ions; Knowledge; Label; Laboratories; Ligands; Membrane; Modeling; Molecular; Narcotic Analgesics; North Carolina; Nuclear; Opiates; Opioid; Opioid Receptor; Overdose; Pain management; Peptides; Pharmacologic Substance; Photons; Physiological; Play; Process; Proteins; Proteome; Protocols documentation; Publications; Reagent; Receptor Signaling; Recording of previous events; Reporting; Research Institute; Resolution; Rest; Role; Signal Transduction; Stimulus; Structure; Structure-Activity Relationship; Substance abuse problem; System; Techniques; Therapeutic; Universities; Work; addiction; chronic pain; design; extracellular; heroin overdose; high risk; high throughput screening; insight; large scale production; member; protocol development; public health relevance; receptor; receptor binding; response; small molecule; transmission process

 DESCRIPTION (provided by applicant): Funds are requested to carry out NMR studies to develop a deeper understanding of the dynamic behavior of G-Protein Coupled Receptors (GPCRs) and their signaling complexes to enable the construction of a better model of their mechanism of action. GPCRs are critical eukaryotic signal transduction gatekeepers which recognize a variety of extracellular stimuli, including photons, ions, small molecules, peptides and proteins. They transmit the resulting extracellular signal across the membrane by coupling to different intracellular proteins which then activate downstream effectors and trigger cascades of cellular and physiological responses. This has profound therapeutic implications, making these receptors the targets of ~30% of currently prescribed drugs. Developing a better understanding of how they work is, therefore, expected to help in developing new and better drugs targeting them. Although there has been a dramatic growth in the number of structures of GPCRs bound to antagonists, agonists including biased ligands, inverse agonists, and allosteric modulators, the molecular mechanisms of activation remain poorly understood. The proposed study rests on the premise that needed clarity can be achieved with a better understanding of the dynamic behavior of the system. We plan to characterize the dynamic behavior of all the members of the opioid family of receptors (Class A, -subfamily). There are four members in this family - the classical opioid receptors (OR) -, -, , (MOR, KOR, DOR) and the nociceptn receptor (NOP), all of which are coupled predominantly to heterotrimeric Gi/Go proteins. ORs are acted on by opioids which have had a long history in both chronic and acute pain control and addiction. This funding request is to support studies on KOR and the development of new knowledge and protocols that could be applied to the rest of the family. We will achieve a better understanding of the mechanism of action including the activation process through three specific aims: (1) Develop an understanding of the dynamic conformational landscape of KOR in the active and inactive state, (2) Develop an understanding of the dynamic behavior of the KOR signaling complex, and (3) Develop an understanding of the KOR-dynorphin interaction in the "low" and "high" affinity states. In this proposal, we will use our newly developed 19F NMR approach to characterize the dynamic behavior of KOR and standard NMR techniques for 2H/13C/15N NMR studies of the peptide dynorphin and the signaling partners to characterize changes to their dynamic behavior on binding to KOR. Studies will allow us to explore various questions, answers to which will help us build a better model describing the activation process. In addition, protocols and reagents generated in the study should help establish a new platform for developing new drugs targeting opioid receptors with reduced or minimal side-effects.

 描述（由申请人提供）：需要资金进行核磁共振研究，以更深入地了解 G 蛋白偶联受体 (GPCR) 及其信号复合物的动态行为，从而能够构建更好的作用机制模型GPCR 是重要的真核信号转导看门人，可识别各种细胞外刺激，包括光子、离子、小分子、肽和蛋白质，它们将产生的细胞外信号跨膜传输。通过与不同的细胞内蛋白质偶联，然后激活下游效应器并触发细胞和生理反应的级联反应，这具有深远的治疗意义，使这些受体成为目前处方药物中约 30% 的靶点。因此，尽管与拮抗剂、激动剂（包括偏向配体、反向激动剂和变构调节剂）结合的 GPCR 结构数量急剧增加，但预计将有助于开发新的、更好的靶向药物。激活的分子机制仍然知之甚少，所提出的研究的前提是可以通过更好地理解系统的动态行为来实现所需的清晰度，我们计划描述阿片受体家族所有成员的动态行为。 （A 类， 亚科）该家族有四个成员 - 经典阿片受体 (OR) -、-、、（MOR、KOR、DOR）和伤害感受器受体。 (NOP)，所有这些都主要与异三聚体 Gi/Go 蛋白偶联，并由阿片类药物作用，阿片类药物在慢性和急性疼痛控制和成瘾方面有着悠久的历史。开发可应用于该家族其他成员的新知识和方案我们将通过三个具体目标更好地理解包括激活过程在内的作用机制：（1）加深对 KOR 动态构象景观的理解。在活动和非活动状态下， (2) 了解 KOR 信号复合物的动态行为，以及 (3) 了解“低”和“高”亲和力状态下的 KOR-强啡肽相互作用。开发了 19F NMR 方法来表征 KOR 的动态行为，并开发了用于肽强啡肽和信号传导伴侣的 2H/13C/15N NMR 研究的标准 NMR 技术，以表征其与结合的动态行为的变化KOR。研究将使我们能够探索各种问题，这些问题的答案将帮助我们建立一个更好的模型来描述激活过程。此外，研究中产生的方案和试剂应有助于建立一个新平台，用于开发针对阿片受体的新药物。减少或最小化副作用。