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（由适用提供）：要求资金进行NMR研究，以更深入地了解G蛋白耦合受体（GPCR）及其信号传导复合物的动态行为，以实现更好的作用机理模型。 GPCR是关键的真核信号转导守门人，识别各种细胞外刺激，包括照片，离子，小分子，肽和蛋白质。它们通过耦合到不同的细胞内蛋白，从而激活下游效应并触发细胞和物理反应的级联反应，从而在整个膜上传输所得的细胞外信号。这具有深刻的理论含义，使这些受体成为当前处方药的30％的靶标。因此，对他们的工作方式有了更好的了解，这是期望有助于开发针对它们的新药物。尽管与拮抗剂结合的GPCR的结构数量有动态增长，但包括偏见的配体，反向激动剂和变构调节剂在内的激动剂，但激活的分子机制仍然很少了解。拟议的研究取决于可以通过更好地理解系统动态行为的前提。我们计划表征阿片受体家族的所有成员（A类，育种）的动态行为。这个家族中有四个成员 - 经典的阿片受体（OR）-，-，，（MOR，KOR，DOR）和NociTeptn受体（NOP），所有这些受体（NOP）主要均与异核酸GI/GO/GO蛋白耦合。 ORS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中都有悠久的历史。这项资金要求是为了支持有关Kor的研究以及可以应用于家庭其他成员的新知识和协议的发展。我们将更好地理解作用机制，包括通过三个特定目的的激活过程：（1）在主动和不活跃状态下对Kor的动态景观有一种理解，（2）（2）对Kor信号传导复合物的动态行为有了了解，（3）对Kor-Dynorphin的相互作用的理解在“低”和“高中”和“高大的”状态。在此提案中，我们将使用新开发的19F NMR方法来表征Kor和标准NMR技术的动态行为，用于2H/13C/15N NMR对肽dynorphin的NMR研究和信号伙伴的研究，以表征其与Kor结合的动态行为的变化。研究将使我们能够探索各种问题，这些问题将有助于我们建立一个描述激活过程的更好模型。此外，研究中产生的方案和试剂应有助于建立一个新的平台，以开发针对阿片类药物受体的新药物，其副作用降低或最小。