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

PACAP/VIP GPCR 亚型的结构、机制和调控

基本信息

批准号：
10001570
负责人：
Jianing Li
金额：
$ 34.41万
依托单位：
UNIVERSITY OF VERMONT & ST AGRIC COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-01 至 2023-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10001570
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 arrestin 2 behavioral response beta-arrestin biological adaptation to stress chronic pain design experimental study feeding glucose metabolism guided inquiry inhibitor/antagonist innovation insight interdisciplinary approach models and simulation molecular dynamics nervous system disorder neurodevelopment novel pituitary adenylate cyclase activating polypeptide polypeptide protein structure receptor secretin receptor simulation small molecule small molecule therapeutics stress related disorder synergism therapeutic development therapeutic target vasoactive intestinal peptide 2 receptor vasoactive intestinal peptide receptor 1

项目摘要

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.

在药理学上重要的 B 类 G 蛋白偶联受体 (GPCR) 中，PAC1/VPAC 垂体腺苷酸环化酶激活多肽（PACAP， ADCYAP1) 和血管活性肠肽 (VIP) 与多种疾病有关，包括慢性疼痛和压力相关的行为异常是全球最普遍的神经系统疾病之一然而，开发针对这些受体的潜在疗法的试剂是当今的挑战。由于缺乏全长 PAC1/VPAC 受体结构信息和机制而受到阻碍了解配体结合如何差异性地驱动受体激活的构象变化因此，该提案的总体目标是采用最先进的技术。计算方法来模拟 PAC1/VPAC 受体结构和动力学，目标是开发调节其功能的小分子化合物对于每种受体亚型，都有独特的识别。允许特定配体相互作用和转导蛋白关联的结构特征将促进为了实现这一目标，小分子配体的合理设计和优化将实现三个具体目标。追求：（1）建模和比较决定神经肽选择性的结构和机制生理相关 PAC1Null、PAC1Hop1、VPAC1 和 VPAC2 受体亚型之间的功能 (2) 描述 PAC1/VPAC 受体-转导蛋白相互作用和特异性；(3) 开发和优化小分子选择性 PAC1 和 VPAC 受体调节我们的初步数据有。建立了 PAC1Null 受体的建模方法，展示了不同的信号传导行为 PAC1/VPAC 受体，并确定了两种不同的 PAC1Null 拮抗剂，在目标 1 和 2 下，我们将使用同源建模、蛋白质结构细化和分子动力学模拟来研究不含配体并与神经肽 (PACAP/VIP) 或转导蛋白复合的受体（Gs/Gq/β-arrestins）特别是，我们将阐明对于配体特异性和关键的结合位点。促进长期信号传导的构象状态将提供信息。和/或证实受体模型在目标 3 下，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，以开发选择性小分子调节剂。特别是，针对正构和变构位点的策略将受到测试。该方法具有创新性，因为它提供了一种无与伦比的全面手段来调查此外，拟议的研究是各种条件和功能状态下的 PAC1/VPAC 受体。意义重大，因为它将弥合理解分子结构和动力学如何进行的基本差距最后，考虑到目前治疗的局限性，这些研究可以决定 PAC1/VPAC 受体机制。可能为治疗具有挑战性的神经系统疾病提供新的机会和方法。