Mechanisms of Adhesion GPCR Action

粘附 GPCR 作用机制

基本信息

批准号：
9352861
负责人：
Gregory Gordon Tall
金额：
$ 31万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-09-15 至 2019-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9352861
关键词：
ADGR1 gene Adhesions Adult Affinity Chromatography Agonist Autistic Disorder Bilateral Binding Biochemical Biological Assay Cell Adhesion Cell Adhesion Molecules Cell membrane Cells Chemicals Cleaved cell Collagen Communities Complex Coupling Data Disease Dissociation Elements Engineering Epitopes Event Extracellular Matrix Extracellular Matrix Proteins Extracellular Space Family Family member G-Protein-Coupled Receptors G-substrate GTP-Binding Proteins Heterotrimeric GTP-Binding Proteins Hormonal Investigation Lead Libraries Ligands Link Luciferases MALDI-TOF Mass Spectrometry Malignant Neoplasms Measurement Measures Mediating Membrane Microgyria Molecular Mutation N-terminal Peptides Photoaffinity Labels Physiological Proteins Receptor Activation Regulation Research Resources Role Side Signal Transduction Signaling Protein Site Small Molecule Chemical Library Specificity Stimulus Testing Time Transmembrane Domain Validation Work cell motility developmental disease extracellular inhibitor/antagonist member mutant new technology prospective protein activation receptor receptor coupling receptor expression receptor function reconstitution response small molecule synthetic peptide therapy development tool transglutaminase 2

ADGR1 gene Adhesions Adult Affinity Chromatography Agonist Autistic Disorder Bilateral Binding Biochemical Biological Assay Cell Adhesion Cell Adhesion Molecules Cell membrane Cells Chemicals Cleaved cell Collagen Communities Complex Coupling Data Disease Dissociation Elements Engineering Epitopes Event Extracellular Matrix Extracellular Matrix Proteins Extracellular Space Family Family member G-Protein-Coupled Receptors G-substrate GTP-Binding Proteins Heterotrimeric GTP-Binding Proteins Hormonal Investigation Lead Libraries Ligands Link Luciferases MALDI-TOF Mass Spectrometry Malignant Neoplasms Measurement Measures Mediating Membrane Microgyria Molecular Mutation N-terminal Peptides Photoaffinity Labels Physiological Proteins Receptor Activation Regulation Research Resources Role Side Signal Transduction Signaling Protein Site Small Molecule Chemical Library Specificity Stimulus Testing Time Transmembrane Domain Validation Work cell motility developmental disease extracellular inhibitor/antagonist member mutant new technology prospective protein activation receptor receptor coupling receptor expression receptor function reconstitution response small molecule synthetic peptide therapy development tool transglutaminase 2

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项目摘要

Summary/Abstract: G protein Coupled Receptors (GPCRs) transduce information from chemical or hormonal stimuli across the cell membrane to activate heterotrimeric G proteins and thereby elicit physiological/cellular responses. Adhesion G Protein Coupled Receptors (aGPCRs) comprise a 33-member sub-family of GPCRs that have become subjects of intense investigation due to an expanding understanding of the roles these receptors have in developmental disorders and adult cancers. Despite the intense study, direct measurements of heterotrimeric G protein activation by any aGPCR family member had never been made until our very recent work. We leveraged a new technology that we developed to purify large quantities of all classes of heterotrimeric G protein α subunits and successfully reconstituted G protein activation mediated by two adhesion GPCRs (GPR56 and GPR110). We determined the G protein subtypes that GPR56 and GPR110 activate and found that the extracellular fragments of both receptors inhibit the G protein activating activity of the receptor portion that is intercalated in the membrane (TM or Trans-membrane domain). Receptor activation occurs when the inhibitory extracellular fragment is dissociated from the TM domain, perhaps by the action of protein ligands in the extracellular space that bind to it. This act unmasks a short portion of the TM domain that acts upon itself in positive regulatory fashion (a tethered agonist). These preliminary data lead us to propose that the entire adhesion GPCR class shares a common mechanism of action in being regulated by tethered agonists. We seek to verify the mechanism for GPR56, GPR114, and the Group VI adhesion GPCRs and perform detailed studies to define the exact composition of the tethered agonist and the way that it engages the TM domain. We will conduct studies to investigate the way that natural ligands of the receptors act to reveal the tethered agonist while we molecularly identify authentic receptor ligands. Screens of small molecule chemical libraries will be conducted to identify synthetic adhesion GPCR modulators. These compounds will greatly aid our mechanistic studies as research tools and may find use as leads to block the pathophysiologic actions of adhesion GPCRs in disease contexts. Understanding the mechanism of action of adhesion GPCRs is a pre- requisite to the development of therapies for the increasing number of diseases ascribed to this GPCR-class. Our work strives to elucidate the mechanism of adhesion GPCR action.

摘要/摘要： G蛋白偶联受体（GPCR）转化了细胞中化学或激素刺激的信息膜激活异三聚体G蛋白，从而引起生理/细胞反应。粘附g 蛋白质偶联受体（AGPCR）构成了已成为GPCR的33个成员的子族由于对这些受体在发育障碍和成人癌症。尽管进行了激烈的研究，但直接测量异三聚体直到我们最近的工作，任何AGPCR家族成员的G蛋白激活都从未发生过。我们利用了我们开发的新技术来净化大量所有类别的异三个类别G 蛋白质α亚基，并成功地重构了由两个粘附GPCR介导的G蛋白激活（GPR56和GPR110）。我们确定了GPR56和GPR110激活的G蛋白亚型的G蛋白亚型并发现两个受体的细胞外片段都抑制了受体部分的G蛋白激活活性在膜（TM或跨膜结构域）中插入。当受体激活发生抑制性细胞外碎片与TM结构域分离，也许是通过蛋白质配体在与之结合的细胞外空间。此行为揭示了TM域的一小部分以积极的监管方式（一种束缚的激动剂）。这些初步数据使我们提出了整个粘附GPCR类具有由束缚激动剂调节的共同作用机理。我们寻求验证GPR56，GPR114和组VI粘合剂GPCR的机制，并进行了详细的作用研究定义束缚激动剂的确切组成及其与TM结构域的参与方式的研究。我们将进行研究以研究接收器的天然配体揭示束缚的方式激动剂时，我们分子鉴定正宗受体配体。小分子化学文库的屏幕将进行识别合成粘合剂GPCR调节剂。这些化合物将极大地帮助我们机械研究作为研究工具，可能会发现用作阻止的铅，以阻止疾病环境中的粘附GPCR。了解粘附GPCR的作用机理是一种前需要开发分配给该GPCR级的疾病数量增加的疗法。我们的工作旨在阐明粘合剂GPCR作用的机制。