CRF and urocortins and their receptors

CRF 和尿皮质素及其受体

• 批准号: 7429657
• 负责人: WYLIE W. VALE
• 金额: $ 54.81万
• 依托单位: SALK INSTITUTE FOR BIOLOGICAL STUDIES
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-06-01 至 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7429657
• 关键词: Address; Adrenal Glands; Affinity; Agonist; Beta Cell; Binding Sites; Biochemical; Biological; C-terminal; CRF receptor type 2; Cardiac Myocytes; Cardiovascular system; Cells; Collaborations; Complex; Corticotropin-Releasing Hormone; Corticotropin-Releasing Hormone Receptors; Development; Disease; Endocrine; Environment; Extracellular Domain; Family; Funding; Genes; Glucagon; Glucose; Grant; Head; Human; Human Cell Line; Institutes; Insulin; Islets of Langerhans; Knockout Mice; Knowledge; Ligand Binding; Ligands; Localized; MAP Kinase Gene; Mediating; Membrane; Metabolic stress; Methods; Molecular; Mus; N-terminal; Nature; Pancreas; Pancreatic Hormones; Peptide Receptor; Peptides; Physiological; Pituitary Gland; Play; Process; Production; Protein Isoforms; Proteins; RNA Interference; RNA Splicing; Recombinants; Regulation; Rodent; Role; Site; Solutions; Staging; Stress; Structure; Tissues; Variant; Vasopressins; Work; base; design; immunoreactivity; insight; insulin secretion; interest; islet; mutant; neutralizing antibody; novel; programs; protein protein interaction; receptor; receptor structure function; research study; response; stem; urocortin

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.

我们正在探讨以下假设：皮质激素释放因子（CRF）/尿道素家族及其同源受体在调节压力和其他情况的适应性反应中起着特定而关键的作用。在以前的授予期内，该项目在整个程序中与CRF进行了特征，克隆了2个CRF受体CRFR1和CRFR2，并确定了对两个受体的亲和力很高的尿素素（UCN）1。在最后一个时期，我们和Sawchenko项目发现了另外两个UCN 2和3的配体，它们对CRFR2具有很高的选择性。我们专注于探索CRF​​R的N末端第一个细胞外域（ECD1）的作用，其中包括肽激动剂和拮抗剂的关键结合位点。我们设计并表达了ECD1的可溶形式 CRFR1和CRFR2BETA保留了适合配体的亲和力，并与Salk Institute的Roland Riek合作，通过NMR解决CRFR2Beta-ECD1的结构。在下一个时期，我们将与R. Riek，Core C和Rivier项目领导的一个新项目紧密合作，以解决CRFR1，CRFR2Alpha和CRFR2Beta的受体ECD1-配体复合物的结构，以及主要由R2alpha的ECD1组成的新型溶解剪接变体。 ECD1中蛋白质蛋白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CRFR2BETA和ERBB2之间看到的功能相互作用可能部分取决于两个受体之间的相互作用。 CRFR2的小鼠无效的结果表明，该受体及其配体在中枢神经系统，心血管系统和胰岛中具有多种作用。缺乏3种尿素蛋白酶的小鼠是与原始Lee项目合作开发的，并且将在该项目中分析垂体/肾上腺和胰腺激素的调节，以及与CRF相关的受体的表达以及剩余的配体的表达。最后，我们将研究UCN 3（及以后的UCN 2）的重要性，显示在β细胞中产生并刺激 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，代谢和与压力相关疾病的见解。