Dynamics of the Opioid Receptor Signaling Complex

阿片受体信号复合物的动力学

• 批准号: 9095376
• 负责人: Kurt Wuthrich
• 金额: $ 38.02万
• 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/9095376
• 关键词: Acute Pain; Address; Adverse effects; Affinity; Agonist; Alcohols; Amphetamines; Behavior; Behavioral; Binding; Biochemistry; Biological Assay; California; Cessation of life; Cocaine; Complex; Coupled; Coupling; Data; Development; Drug Prescriptions; Drug Targeting; Dynorphins; Engineering; Evaluation; Family; Formulation; France; Funding; Funding Applicant; Future; G-Protein-Coupled Receptors; GPR6 gene; GTP-Binding Proteins; Gatekeeping; Goals; Growth; Health; 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; beta-2 Adrenergic Receptors; chronic pain; design; extracellular; heroin overdose; high risk; high throughput screening; insight; large scale production; member; new therapeutic target; protocol development; 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）：要求提供资金进行NMR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是真核生物信号转导的重要看门人，它识别各种细胞外刺激，包括光子、离子、小分子、肽和蛋白质。它们通过偶联到不同的细胞内蛋白质，然后激活下游效应器并触发细胞和生理反应的级联反应，从而跨膜传递所得的细胞外信号。这具有深远的治疗意义，使这些受体成为目前约30%处方药物的靶点。因此，更好地了解它们是如何工作的，预计将有助于开发针对它们的新的和更好的药物。尽管与拮抗剂、激动剂（包括偏向性配体、反向激动剂和变构调节剂）结合的GPCR的结构数量急剧增长，但激活的分子机制仍然知之甚少。所提出的研究基于这样的前提，即通过更好地理解系统的动态行为可以实现所需的清晰度。我们计划描述阿片受体家族（A类，β-亚家族）所有成员的动态行为。该家族中有四个成员-经典阿片受体（OR）β-、（莫尔、KOR、DOR）和伤害感受素受体（NOP），所有这些受体主要与异源三聚体Gi/Go蛋白偶联。阿片类药物对OR起作用，阿片类药物在慢性和急性疼痛控制和成瘾方面都有悠久的历史。这项资金申请是为了支持对KOR的研究，以及开发可适用于该家族其他成员的新知识和协议。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）了解KOR在活性和非活性状态下的动态构象景观，（2）了解KOR信号复合物的动态行为，（3）了解KOR-dynorphin在“低”和“高”亲和力状态下的相互作用。在这项提案中，我们将使用我们新开发的19 F NMR方法来表征KOR的动态行为，并使用标准NMR技术对肽强啡肽和信号伴侣进行2 H/13 C/15 N NMR研究，以表征其与KOR结合时动态行为的变化。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该有助于建立一个新的平台，用于开发靶向阿片受体的新药，减少或减少副作用。