Structure, Mechanism, and Regulation of PACAP/VIP GPCR subtypes

PACAP/VIP GPCR 亚型的结构、机制和调控

• 批准号: 10473545
• 负责人: Jianing Li
• 金额: --
• 依托单位: UNIVERSITY OF VERMONT & ST AGRIC COLLEGE
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-01 至 2022-09-02
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10473545
• 关键词: Adenylate Cyclase; Agonist; Allosteric Site; Amino Acid Receptors; Anxiety; Arrestins; Attention; Behavior; Behavioral; Binding; Binding Sites; Biochemical; Biological Assay; Calcium; Cell physiology; Cellular Assay; Chronic stress; Circadian Rhythms; Complex; Computing Methodologies; Coupling; Data; Development; Disease; Docking; Endocrine; Equilibrium; Face; Family; Free Energy; G-Protein-Coupled Receptors; GTP-Binding Proteins; Glucagon; Goals; Heterotrimeric GTP-Binding Proteins; Homeostasis; Homology Modeling; Hypercalcemia; Immune; In Vitro; Inflammation; Knowledge; Lead; Length; Ligand Binding; Ligands; Mediating; Mental Depression; Metabolic; Metabolic Diseases; Methodology; Migraine; Mission; Modeling; Molecular; Molecular Conformation; Molecular Structure; Mutagenesis; Nervous system structure; Neurologic; Neuropeptides; Nociception; Non-Insulin-Dependent Diabetes Mellitus; Obesity; Organic Synthesis; Osteoporosis; Pathway interactions; Peptide Receptor; Peptides; Pharmacology; Physiologic Thermoregulation; Physiological; Pituitary Gland; Proteins; Public Health; Reagent; Receptor Activation; Receptor Signaling; Regulation; Research; Role; Second Messenger Systems; Signal Transduction; Signaling Molecule; Specificity; Stress; Structure; System; Technology; Testing; Therapeutic; Therapeutic Studies; Time; Transducers; United States National Institutes of Health; VIPR1 gene; Variant; Vasoactive Intestinal Peptide; Water; Weight Gain; Work; antagonist; arrestin 2; behavioral response; beta-arrestin; biological adaptation to stress; chronic pain; experimental study; feeding; glucose metabolism; guided inquiry; inhibitor; innovation; insight; interdisciplinary approach; models and simulation; molecular dynamics; nervous system disorder; neurodevelopment; novel; pituitary adenylate cyclase activating polypeptide; polypeptide; protein structure; rational design; receptor; secretin receptor; simulation; small molecule; small molecule therapeutics; stress related disorder; synergism; therapeutic development; therapeutic target; vasoactive intestinal peptide 2 receptor

Among the pharmacologically important Class B G protein-coupled receptors (GPCRs), the PAC1/VPAC receptors (ADCYAP1R1/VIPR1/VIPR2) for pituitary adenylate cyclase activated polypeptide (PACAP, ADCYAP1) and vasoactive intestinal peptide (VIP) have been implicated in several disorders, including chronic pain and stress-related behavioral abnormalities which are among the most prevalent global neurological challenges today. However, the development of reagents targeting these receptors for potential therapeutics has been hampered by the lack of full-length PAC1/VPAC receptor structural information, and mechanistic understandings of how ligand binding can differentially drive conformational changes for receptor activation and biased signaling. Accordingly, the overarching aim of this proposal is to employ state-of-the-art computational methods to model PAC1/VPAC receptor structure and dynamics, with the goal of developing small-molecule compounds to modulate their functions. For each receptor subtype, the identification of unique structural features that allow for specific ligand interactions and transducer protein associations will facilitate the rational design and optimization of small-molecule ligands. Towards that goal, three specific aims will be pursued: (1) to model and compare structures and mechanisms that determine neuropeptide selectivity and function among the physiologically relevant PAC1Null, PAC1Hop1, VPAC1, and VPAC2 receptor subtypes; (2) to delineate PAC1/VPAC receptor-transducer protein interactions and specificity; and (3) to develop and optimize small molecules for selective PAC1 and VPAC receptor regulation. Our preliminary data have established the modeling methodology with the PAC1Null receptor, demonstrated distinct signaling behaviors of the PAC1/VPAC receptors, and identified two different PAC1Null antagonists. Under Aims 1 and 2, we will use homology modeling, protein structure refinement, and molecular dynamics simulations to study the receptors that are ligand-free and complexed with a neuropeptide (PACAP/VIP) or a transducer protein (Gs/Gq/β-arrestins). In particular, we will elucidate the binding sites that are key for ligand specificity as well as conformational states that facilitate long-term signaling. Receptor mutagenesis and constructs will inform and/or substantiate the receptor models. Under Aim 3, we will integrate molecular docking and simulations, organic synthesis, and molecular and cellular assays to develop selective small-molecule modulators. Especially, the strategies to target the orthosteric and allosteric sites will be tested. Our multidisciplinary approach is innovative as it provides an unparalleled and comprehensive means to investigate the PAC1/VPAC receptors in various conditions and functional states. Further, the proposed research is significant, because it will close fundamental gaps in understanding how molecular structures and dynamics can dictate PAC1/VPAC receptor mechanisms. Finally, given current limitations in therapeutics, these studies may offer new opportunities and approaches to treat challenging neurological disorders.

在具有药理学重要的B级蛋白组合受体（GPCR）中，PAC1/VPAC 垂体腺苷酸环化酶活化多肽的受体（ADCYAP1R1/VIPR1/VIPR2）（PACAP，PACAP，，PACAP， ADCYAP1）和血管活性肠肽（VIP）已与慢性 疼痛和与压力相关的行为异常是最普遍的全球神经学之一 但是，今天的挑战。 由于缺乏全长PAC1/VPAC受体结构信息和机械性能所阻碍 对配体结合如何差异化驱动置换变化以使受体激活的理解 和有偏见的信号。 建模PAC1/VPAC受体结构和动力学的计算方法，目的是开发 小分子化合物可调节其功能。 允许特定配体相互作用和传感器蛋白缔合的结构特征将有助于 小分子配体的合理设计和优化。 追求：（1）建模和比较确定神经肽选择性和的结构和机制 在硫代基元相关的PAC1NULL，PAC1HOP1，VPAC1和VPAC2受体亚型之间的功能；（2） 描绘PAC1/VPAC受体 - 转化蛋白的相互作用和（3） 优化针对选择性PAC1和VPAC受体调控的小分子。 用PAC1NULL受体建立了建模方法，表现出不同的信号行为 在PAC1/VPAC受体中，在AIM 1和2下鉴定了两个不同的PAC1NULL拮抗剂。 使用同源性建模，蛋白质结构的细化和分子动力学模拟来研究您 不含配体并与神经肽（PACAP/VIP）或换能蛋白的受体竞争 （尤其是GS/GQ/β-Arresstins）。 促进长期信号传导的构象状态。 和/或在AIM 3下证实受体模型，我们将整合分子对接和模拟， 有机合成以及分子和细胞测定，以开发选择性的小分子调节剂。 尤其是，将测试到正构和同层站点的策略。 方法具有创新性，因为它提供了一种无与伦比的全面手段来调查您 PAC1/VPAC受体在各种条件和功能状态下。 重要的是，因为它将在理解分子结构和动力学方面截断基本差距 可以决定PAC1/VPAC受体机制。 可能会提供新的机会和方法来治疗具有挑战性的神经局部疾病。