RAMP-altered Class B GPCR Hormone Recognition

RAMP 改变的 B 类 GPCR 激素识别

• 批准号: 8422162
• 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
• 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/8422162
• 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 Delivery Systems; Drug Design; 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

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. PUBLIC HEALTH RELEVANCE: This project addresses how peptide hormones bind to receptors on the surface of human cells. The hormones we study are important regulators of the cardiovascular system and blood glucose levels. The receptors are members of a large family of receptors known as GPCRs that regulate virtually all aspects of human physiology. Discovering how these hormones bind to their receptors is important because it will aid the development of drugs that target the receptors for the treatment of cardiovascular disorders and diabetes. These studies are also important for the general knowledge we will gain about GPCRs, which will be broadly applicable to several areas of human health and disease.

描述(申请人提供)：人降钙素(CTR)和类降钙素(CLR)受体是细胞表面B类G蛋白偶联受体(GPCRs)，与受体活性修饰蛋白(RAMP)共受体结合，通过降钙素家族多肽激素介导信号传递。肾上腺髓质素(AM)、降钙素基因相关肽(CGRP)和胰淀素(AMY)是通过与由CLR或CTR和三个RAMP之一组成的特定分子复合体结合而产生的不同的生物学作用。AM和CGRP分别通过CLR-RAMP2或3和CLR-RAMP1复合体调节心血管系统。AMY通过与RAMP1、2或3相关的CTR信号来调节血糖水平。CLR/CTR-RAMP复合体是重要的药物靶点。针对CLR-RAMP复合体的治疗剂有可能治疗急性心肌梗死、肺动脉高压、偏头痛、癌症和其他几种疾病。CTR-RAMP复合体是糖尿病药物SymlinTM的靶点。尽管靶向CLR/CTR-RAMP复合体具有临床价值，但对于多肽激素识别和调节CLR/CTR激素特异性的RAMP的分子机制尚不清楚，这在很大程度上是因为缺乏多肽-受体复合体的结构信息。激素结合的亲和力和特异性在很大程度上由CLR、CTR和RAMP整合膜蛋白的胞外区(ECD)决定。该建议的目标是确定AM、CGRP和AMY与其各自的CLR/CTR ECD-RAMP ECD复合体结合的晶体结构，以便定义决定选择性多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联系起来。我们提出了以下三个目标：(1)确定人CLR-RAMP2 ECD复合体识别AM的机制。(2)确定RAMP1如何赋予CLR-RAMP1 ECD复合体CGRP选择性。(3)确定人CTR-RAMP1ECD复合体识别AMY的机制。实现这些目标将确定RAMP改变的B类GPCR激素识别的分子基础，并为指导针对受体的治疗药物的合理设计提供结构模板。 与公共健康相关：该项目解决了多肽激素如何与人类细胞表面受体结合的问题。我们研究的荷尔蒙是心血管系统和血糖水平的重要调节器。这些受体是被称为GPCRs的受体大家族的成员，GPCRs几乎调节人类生理的所有方面。发现这些激素如何与它们的受体结合是很重要的，因为这将有助于开发针对受体的药物，用于治疗心血管疾病和糖尿病。这些研究对于我们将获得的关于GPCRs的一般性知识也很重要，这些知识将广泛适用于人类健康和疾病的几个领域。