Secretin Receptor Structure, Function, and Regulation

促胰液素受体的结构、功能和调节

基本信息

批准号：
7578221
负责人：
LAURENCE J MILLER
金额：
$ 36.48万
依托单位：
MAYO CLINIC ARIZONA
依托单位国家：
美国
项目类别：
财政年份：
2005
资助国家：
美国
起止时间：
2005-02-01 至 2010-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7578221
关键词：
Adrenergic Receptor Agonist Amino Acids Area Attention Automobile Driving Binding Biochemical Biological Bioluminescence Cell membrane Charge Complex Cysteine DNA Sequence Rearrangement Data Development Disease Dominant-Negative Mutation Drug Delivery Systems Energy Transfer Environment Epitopes Exons Face Family Fluorescence Fluorescence Resonance Energy Transfer Funding G-Protein-Coupled Receptors Generations Glycoproteins Goals Grant Growth Inhibitors Health Heterotrimeric GTP-Binding Proteins Ligand Binding Ligands Malignant Neoplasms Malignant neoplasm of pancreas Measures Modeling Molecular Molecular Conformation Molecular Models Mutagenesis Neoplasms Pattern Peptides Pharmaceutical Preparations Phenotype Phosphorylation Photoaffinity Labels Photons Physiology Play Post-Translational Protein Processing Process Publishing RNA Splicing Reagent Receptor Activation Receptor Signaling Regulation Research Personnel Rhodopsin Role Secretin Series Signal Transduction Site Structure Techniques Testing Variant Viral Work analog base cancer cell cell growth cell growth regulation design disulfide bond drug development inhibitor/antagonist insight member molecular modeling mutant novel novel therapeutics particle pharmacophore programs receptor receptor binding receptor function receptor structure function research study secretin receptor synthetic peptide

项目摘要

DESCRIPTION (provided by applicant): The secretin receptor is prototypic of the important Class B family of G protein-coupled receptors. The long-term goal of this work is to better understand the structure, function, and regulation of this group of receptors, gaining insights that will facilitate the development of new therapeutic strategies and new drugs that can act at these targets. The projects are designed to test, extend, and refine the recently proposed molecular model of the natural agonist-occupied secretin receptor and to elucidate the molecular basis of receptor activation and receptor regulation by oligomerization within the plasma membrane. There are three broad aims for this proposal. The first aim is designed to explore the hypothesis that the amino-terminal domain of the secretin receptor provides a critical ligand-binding pocket that undergoes a conformational rearrangement upon binding the natural peptide agonist. This will be investigated by photoaffinity labeling specific sites within the receptor using series of agonist and antagonist probes, by developing and applying fluorescent indicators within agonist and antagonist probes, and by the application of fluorescence resonance energy transfer techniques. The second aim is designed to explore the molecular mechanism of transduction of the activation signal from the receptor amino terminus to the receptor body, examining the novel hypothesis predicting the presence of an endogenous agonist within the receptor sequence that is exposed upon agonist binding. This will be examined by photoaffinity labeling with charge-modified ligands, site-directed receptor mutagenesis, and biological activity studies using synthetic candidate molecules. The third aim is designed to examine the molecular basis and functional importance of secretin receptor oligomerization as a mechanism to regulate the secretin receptor in health and disease. This will be examined using bioluminescence resonance energy transfer with modified receptor constructs, studying impact on function and receptor association. In addition to wild type receptor, a misspliced variant of the secretin receptor recently described in various neoplasms that has dominant negative inhibitory activity will also be studied. Together, these efforts should provide the finest level of molecular detail available for understanding the structure and mechanisms of ligand binding, activation, and regulation of any receptor in this receptor family.

描述（由申请人提供）：Sectionin受体是G蛋白偶联受体的重要B家族的原型。这项工作的长期目标是更好地了解这组受体的结构，功能和调节，获得见解，以促进新的治疗策略和可以在这些目标上起作用的新药物。这些项目旨在测试，扩展和完善自然占地的秘密蛋白受体的最近提出的分子模型，并阐明质膜内寡聚的受体激活和受体调节的分子基础。该提案有三个广泛的目标。第一个目的旨在探讨以下假设：Sectionin受体的氨基末端结构域提供了一个关键的配体结合口袋，该口袋在结合天然肽激动剂时会经历构象重排。通过在受体中使用一系列激动剂和拮抗剂探针在受体中的特定位点，通过在激动剂和拮抗剂探针中开发和应用荧光指标，以及应用荧光共振能量传递技术，对此进行了调查。第二个目的旨在探索从受体氨基末端到受体体的激活信号转导的分子机制，研究了新的假设，预测了受体序列中内源性激动剂的存在，而受体序列中暴露于激动剂结合的受体序列中。这将通过使用合成候选分子的光相位标记，带电荷修饰的配体，定位的受体诱变和生物学活性研究来检查这一点。第三个目的旨在检验分子基础和功能重要性，作为调节健康和疾病中秘密蛋白受体的一种机制。这将使用具有改良的受体构建体的生物发光共振能量转移来研究，从而研究对功能和受体关联的影响。除了野生型受体外，还将研究在各种肿瘤受体中的错位变体，在各种肿瘤中描述的具有主要的负阴性抑制活性的变体。总之，这些努力应提供最优质的分子细节，以了解该受体家族中任何受体的配体结合，激活和调节的结构和机制。