RAMP-altered Class B GPCR Hormone Recognition

RAMP 改变的 B 类 GPCR 激素识别

基本信息

批准号：
8731948
负责人：
Augen A Pioszak
金额：
$ 28.12万
依托单位：
UNIVERSITY OF OKLAHOMA HLTH SCIENCES CTR
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-09-26 至 2017-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8731948
关键词：
Acute myocardial infarction Address Affinity Agonist Architecture Area Binding Biochemical Biological Biological Assay Blood Glucose Calcitonin Calcitonin Gene-Related Peptide Calcitonin-Gene Related Peptide Receptor Cardiovascular Diseases Cardiovascular system Cell Surface Receptors Cell membrane Cells Chimeric Proteins Clinical Clinical Trials Complex Crystallization Development Diabetes Mellitus Disease Drug Design Drug Targeting Extracellular Domain Family G-Protein-Coupled Receptors Goals Health Hormone Receptor Hormones Human Human calcitonin Integral Membrane Protein Knowledge Link Malignant Neoplasms Mammalian Cell Mediating Methodology Migraine Molecular N-terminal Peptide Receptor Peptides Pharmaceutical Preparations Pharmacology Physiological Physiology Production Proteins Pulmonary Hypertension RAMP1 RAMP2 Recombinants Resolution Signal Transduction Specificity Structure Surface Testing Therapeutic Agents Transmembrane Domain Vasodilator Agents Yeasts adrenomedullin adrenomedullin receptor amylin receptor analog base design experience glycosylation improved islet amyloid polypeptide member next generation peptide hormone protein complex receptor receptor-activity-modifying protein research study structural biology

项目摘要

DESCRIPTION (provided by applicant): The human calcitonin (CTR) and calcitonin-like (CLR) receptors are cell-surface class B G protein-coupled receptors (GPCRs) that associate with receptor activity modifying protein (RAMP) co-receptors to mediate signaling by calcitonin family peptide hormones. The distinct biological actions of the peptides adrenomedullin (AM), calcitonin gene-related peptide (CGRP), and amylin (AMY) are elicited by their binding to specific molecular complexes comprised of CLR or CTR and one of three RAMPs. AM and CGRP regulate the cardiovascular system by signaling through CLR-RAMP2 or 3 and CLR-RAMP1 complexes, respectively. AMY regulates blood glucose levels by signaling through CTR associated with RAMP1, 2, or 3. CLR/CTR-RAMP complexes are important drug targets. Therapeutic agents targeting CLR-RAMP complexes have the potential to treat acute myocardial infarction, pulmonary hypertension, migraine headache, cancer, and several other disorders. CTR-RAMP complexes are targets of the diabetes drug SymlinTM. Despite the clinical value of targeting CLR/CTR-RAMP complexes, the molecular mechanisms of peptide hormone recognition and RAMP modulation of CLR/CTR hormone specificity are poorly understood, in large part because of a lack of structural information for the peptide-receptor complexes. Hormone binding affinity and specificity are largely determined by the extracellular domains (ECDs) of the CLR, CTR, and RAMP integral membrane proteins. The goals of this proposal are to determine crystal structures of AM, CGRP, and AMY bound to their respective CLR/CTR ECD-RAMP ECD complexes in order to define the precise molecular architectures that determine selective peptide binding. In addition, we will correlate structure and function through biochemical and cell-based pharmacological experiments. We propose the following three aims: (1) Determine the mechanism of AM recognition by the human CLR-RAMP2 ECD complex. (2) Determine how RAMP1 confers CGRP selectivity to the CLR-RAMP1 ECD complex. (3) Determine the mechanism of AMY recognition by the human CTR-RAMP1 ECD complex. Achieving these aims will define the molecular bases for RAMP-altered class B GPCR hormone recognition and provide structural templates to guide the rational design of therapeutic agents targeting the receptors.

描述（由申请人提供）：人降钙素 (CTR) 和降钙素样 (CLR) 受体是细胞表面 B 类 G 蛋白偶联受体 (GPCR)，与受体活性修饰蛋白 (RAMP) 共受体结合以介导信号传导通过降钙素家族肽激素。肾上腺髓质素 (AM)、降钙素基因相关肽 (CGRP) 和胰岛淀粉样多肽 (AMY) 肽的独特生物作用是通过它们与由 CLR 或 CTR 以及三个 RAMP 之一组成的特定分子复合物的结合而引起的。 AM 和 CGRP 分别通过 CLR-RAMP2 或 3 和 CLR-RAMP1 复合物发出信号来调节心血管系统。 AMY 通过与 RAMP1、2 或 3 相关的 CTR 发出信号来调节血糖水平。CLR/CTR-RAMP 复合物是重要的药物靶点。针对 CLR-RAMP 复合物的治疗药物具有治疗急性心肌梗塞、肺动脉高压、偏头痛、癌症和其他几种疾病的潜力。 CTR-RAMP 复合物是糖尿病药物 SymlinTM 的靶标。尽管靶向 CLR/CTR-RAMP 复合物具有临床价值，但肽激素识别和 CLR/CTR 激素特异性的 RAMP 调节的分子机制仍知之甚少，很大程度上是因为缺乏肽-受体复合物的结构信息。激素结合亲和力和特异性很大程度上取决于 CLR、CTR 和 RAMP 整合膜蛋白的胞外结构域 (ECD)。该提案的目标是确定 AM、CGRP 和 AMY 与其各自的 CLR/CTR ECD-RAMP ECD 复合物结合的晶体结构，以便定义确定选择性肽结合的精确分子结构。此外，我们将通过生化和基于细胞的药理学实验将结构和功能关联起来。我们提出以下三个目标：（1）确定人类CLR-RAMP2 ECD复合物识别AM的机制。 (2) 确定 RAMP1 如何赋予 CLR-RAMP1 ECD 复合物 CGRP 选择性。 (3)确定人CTR-RAMP1 ECD复合物识别AMY的机制。实现这些目标将定义 RAMP 改变的 B 类 GPCR 激素识别的分子基础，并提供结构模板来指导针对受体的治疗药物的合理设计。