Mechanism of activation of natriuretic peptide receptors

利尿钠肽受体的激活机制

• 批准号: 7092182
• 负责人: FOCCO VAN DEN AKKER
• 金额: $ 30.17万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-08-01 至 2009-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7092182
• 关键词: X ray crystallography; atrial natriuretic peptide; biological signal transduction; cell line; cryoelectron microscopy; electron microscopy; enzyme induction /repression; gene mutation; guanylate cyclase; molecular biology; nucleic acid sequence; phosphorylation; polymerase chain reaction; protein purification; receptor expression; structural biology; western blottings

DESCRIPTION (provided by applicant): The atrial natriuretic peptide (ANP) receptor and the homologous c-type natriuretic peptide receptor are homologous yet initiate distinct physiological processes. The ANP receptor is involved in renal and cardiac functions; its activation leads to an increase in natriuresis and diuresis in the kidney and decreased blood pressure. The ANP receptor activating peptide BNP has clinical benefits for congestive heart failure patients. The C-type natriuretic peptide receptor has recently been identified by us as the target for human mutations that cause impaired skeletal growth. Both receptors belong to the family of membrane bound guanylyl cyclases and contain an extracellular ligand binding domain, a transmembrane helix, a kinase-homology domain, a coiled-coil, and guanylyl cyclase domain. Extracellular hormone binding to these receptors leads to the intracellular production of the second messenger cGMP. We have previously determined the 2.0 Angstrom resolution crystal structure of the extracellular ligand binding domain of the ANP receptor. The proposed studies are a continuation of our multi-disciplinary efforts to investigate the mechanism of activation of these receptors. Our first Specific Aim is to investigate whether 1 of the roles of the kinase homology domain is to keep the extracellular domains apart as part of the activation mechanism. This will be tested using structural biology methods with both the intracellular domains as well as with the full length ANP receptor. The second Aim is to test the hypothesis that the domains of the ANP receptor are correctly positioned, and optimized for length. This will be probed by insertional and deletion mutagenesis studies targeting the inter-domain regions. The third Specific Aim is to determine whether the transmembrane helices are involved in lateral movements within the membrane during the activation mechanism. The fourth Aim is to test the hypothesis that a number of the genetic defects in the c-type natriuretic peptide receptor involve mutations that affect the signal transduction mechanism of these receptors. These proposed studies on natriuretic peptide receptors will increase our understanding of mechanisms of activation and function, and could enhance the development of new renal, cardiovascular, or skeletal growth stimulatory drugs.

描述（由申请人提供）：心房利尿肽（ANP）受体和同源的C型纳二肽受体受体是同源的，但启动了不同的生理过程。 ANP受体参与肾功能和心脏功能。它的激活导致肾脏中纳地尿和利尿剂的增加并降低血压。 ANP受体激活肽BNP对充血性心力衰竭患者具有临床益处。我们最近将c型纳特里尿肽受体鉴定为引起骨骼生长受损的人类突变的靶标。这两个受体都属于膜结合的鸟叶烯基环酶家族，并包含一个细胞外配体结合结构域，跨膜螺旋，激酶 - 单学结构域，盘绕型金和鸟叶兰氏循环酶结构域。细胞外激素与这些受体的结合导致第二信使CGMP的细胞内产生。我们先前已经确定了ANP受体的细胞外配体结合结构域的2.0 Angstrom分辨率晶体结构。拟议的研究是我们多学科的努力继续研究这些受体激活机制。我们的第一个具体目的是研究激酶同源性领域的1个作用是将细胞外域与激活机制的一部分保持隔离。这将使用具有细胞内结构域以及全长ANP受体的结构生物学方法进行测试。第二个目的是检验假设ANP受体的域的定位并进行长度优化的假设。这将通过针对域间区域的插入和缺失诱变研究来探讨。第三个具体目的是确定激活机理期间跨膜螺旋是否参与膜内的横向运动。第四个目的是检验以下假设：c型纳替肽受体中的许多遗传缺陷涉及影响这些受体信号转导机制的突变。这些关于亚替肽受体的研究将增加我们对激活和功能机制的理解，并可以增强新的肾脏，心血管或骨骼生长刺激药物的发展。