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：
9128652
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/9128652
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 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 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 targeted treatment

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 G蛋白偶联受体（GPCR），它们与受体活性修饰蛋白（RAMP）共同受体介导信号传导通过降钙素家族肽激素。肽肾上腺甲蛋白酶（AM），降钙素基因相关肽（CGRP）和淀粉蛋白（AMY）的不同生物学作用是通过与CLR或CTR和三个斜坡之一的特定分子复合物结合而引起的。 AM和CGRP分别通过CLR-RAMP2或3和CLR-RAMP1复合物来调节心血管系统。 Amy通过通过与RAMP1、2或3相关的CTR信号传导来调节血糖水平。CLR/CTR-RAMP复合物是重要的药物靶标。靶向CLR-RAMP复合物的治疗剂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共生的靶标。尽管靶向CLR/CTR-RAMP复合物的临床价值，但肽激素识别的分子机制和CLR/CTR激素特异性的坡道调节的理解很少，这在很大程度上是由于缺乏肽受体复合物的结构信息。激素结合亲和力和特异性在很大程度上取决于CLR，CTR和坡道积分膜蛋白的细胞外域（ECD）。该建议的目标是确定与它们各自的CLR/CTR ECD-RAMP ECD复合物结合的AM，CGRP和AMY的晶体结构，以定义确定选择性肽结合的精确分子体系结构。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相关联。我们提出以下三个目的：（1）确定人类CLR-RAMP2 ECD复合物的AM识别机理。（2）确定RAMP1如何赋予CGRP选择性对CLR-RAMP1 ECD复合物。（3）确定人CTR-RAMP1 ECD复合物的AMY识别机制。实现这些目标将定义用于改变坡道的B类GPCR激素识别的分子碱基，并提供结构模板，以指导针对受体的治疗剂的合理设计。