CRF and urocortins and their receptors

CRF 和尿皮质素及其受体

• 批准号: 6956156
• 负责人: WYLIE W. VALE
• 金额: $ 55.19万
• 依托单位: SALK INSTITUTE FOR BIOLOGICAL STUDIES
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-06-01 至 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6956156
• 关键词: binding sites; corticotropin releasing factor; glucagon; hormone receptor; hormone regulation /control mechanism; insulin; laboratory mouse; laboratory rat; pancreatic islet function; pituitary adrenal axis; protein structure function; receptor binding; receptor sensitivity

We are exploring the hypothesis that the Corticotropin Releasing Factor (CRF)/urocortin family of ligands and their cognate receptors play specific and critical roles in the modulation of adaptive responses to stress and other circumstances. In previous granting periods, this Project, with collaborators throughout the Program, characterized CRF, cloned 2 CRF receptors, CRFR1 and CRFR2 and identified urocortin (Ucn) 1, which had high affinity for both receptors. During the last period, we and the Sawchenko Project have found two additional ligands, Ucn 2 and 3, which are highly selective for CRFR2. We have focused on exploring the roles of the N-terminal, first extracellular domain (ECD1) of the CRFRs, which include critical binding sites for peptide agonists and antagonists. We have designed and expressed soluble forms of the ECD1's of CRFR1 and CRFR2beta with retained affinity for appropriate ligands and collaborated with Roland Riek, Salk Institute, to solve the structure of the CRFR2beta-ECD1 by NMR. In the next period, we will work closely with a new project headed by R. Riek, Core C and the Rivier Project to solve the structures of receptor ECD1- ligand complexes for CRFR1, CRFR2alpha and CRFR2beta as well as a novel soluble splice variant comprising mainly the ECD1 of R2alpha. The identification of a protein-protein interaction motif in the ECD1 spawned the testable hypothesis that the functional interaction that we had seen between CRFR2beta and erbB2 in cardiomyocytes may in part depend upon an interaction between the two receptors. Results with mice null for CRFR2 have suggested a variety of roles for this receptor and its ligands in the CNS, cardiovascular system and pancreatic islets. Mice deficient in each of the 3 urocortins have been developed in collaboration with the original Lee project and will be analyzed in this project with respect to the regulation of pituitary/adrenal and pancreatic hormones as well as the expression of CRF-related receptors and remaining ligands. Finally, we will examine the significance of Ucn 3 (and later, Ucn 2), shown to be produced in beta cells and to stimulate insulin and glucagon secretion, as local regulators of islet functions. In the context of the Program, progress in this Project should continue to yield information concerning the molecular nature, regulation and physiologic roles of ligands and receptors of the CRF family and provide insight regarding endocrine, metabolic and stress-related diseases.

我们正在探索的假设，促肾上腺皮质激素释放因子（CRF）/urocortin家族的配体和它们的同源受体在调节适应性反应的压力和其他情况下发挥特定的和关键的作用。在以前的资助期内，本项目与整个项目的合作者一起对CRF进行了表征，克隆了2种CRF受体CRFR 1和CRFR 2，并鉴定了对这两种受体都具有高亲和力的urocortin（Ucn）1。在过去的一段时间里，我们和Sawchenko项目发现了两个额外的配体，Ucn 2和3，它们对CRFR 2具有高度选择性。我们一直专注于探索的N-末端，第一胞外结构域（ECD 1）的CRFR，其中包括关键的肽激动剂和拮抗剂的结合位点的作用。我们已经设计并表达了ECD 1的可溶形式， CRFR 1和CRFR 2 β对适当的配体保持亲和性，并与索尔克研究所的罗兰里克合作，通过NMR解析CRFR 2 β-ECD 1的结构。在接下来的一段时间里，我们将与R。Riek，Core C和Rivier Project解决了CRFR 1、CRFR 2 α和CRFR 2 β的受体ECD 1-配体复合物的结构以及主要包含R2 α的ECD 1的新型可溶性剪接变体。在ECD 1中蛋白质-蛋白质相互作用基序的鉴定产生了可检验的假设，即我们在心肌细胞中观察到的CRFR 2 β和erbB 2之间的功能性相互作用可能部分取决于两种受体之间的相互作用。小鼠CRFR 2缺失的结果表明，该受体及其配体在CNS、心血管系统和胰岛中具有多种作用。与最初的Lee项目合作开发了3种尿皮质素中每一种都缺乏的小鼠，并将在该项目中分析垂体/肾上腺和胰腺激素的调节以及CRF相关受体和剩余配体的表达。最后，我们将研究Ucn 3（以及后来的Ucn 2）的意义，它被证明是在β细胞中产生的， 胰岛素和胰高血糖素分泌，作为胰岛功能的局部调节剂。在该计划的背景下，该项目的进展应继续产生有关CRF家族配体和受体的分子性质、调节和生理作用的信息，并提供有关内分泌、代谢和应激相关疾病的见解。