Natural ligands for 2 endothelin-like orphan receptors

2 个内皮素样孤儿受体的天然配体

• 批准号: 6772433
• 负责人: RAINER K REINSCHEID
• 金额: $ 14.39万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-07-07 至 2005-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6772433
• 关键词: CHO cells; G protein; brain; cell line; endothelin; high performance liquid chromatography; immunocytochemistry; laboratory mouse; ligands; protein sequence; protein structure function; receptor; receptor coupling; tissue /cell culture

DESCRIPTION (provided by applicant): G protein-coupled receptors (GPCRs) form the largest gene family in the human genome and are the most successful biochemical targets of pharmaceutical drugs. For about 140 GPCRs the natural ligands are unknown and they are thus called "orphan" receptors. Identification of these natural ligands is the crucial step to start studying their function and holds the potential for development of novel therapeutic drugs. We have developed a strategy to isolate such natural ligands by expressing the cloned receptors in suitable cell lines and monitoring activation of the receptors after adding fractions of tissue extracts. This strategy will ultimately allow us to purify the active molecules and determine their structure. In this application, we propose to isolate, identify and characterize pharmacologically the natural ligands of two such orphan GPCRs which are structurally related to endothelin receptors. Following the isolation of the natural ligands, we will characterize their pharmacological effects, study their biosynthesis by cloning the precursor proteins that encode them and analyze their distribution in the brain and peripheral organs. These receptors do not bind the known endothelin peptides but have been conserved during evolution across various species, showing that they must serve an important function. Both receptors are highly expressed in the brain and one of them was recently identified as a substrate for parkin, the gene product which is mutated in an autosomal recessive form of Parkinson's disease. Identification of the natural ligands will enable us to study the physiological functions of these two orphan GPCRs under normal and pathological conditions.

描述（由申请人提供）：G蛋白偶联受体（GPCR）构成了人类基因组中最大的基因家族，并且是药物最成功的生化靶标。对于大约140个GPCR，天然配体未知，因此被称为“孤儿”受体。这些天然配体的识别是开始研究其功能并具有开发新型治疗药物的潜力的关键步骤。我们已经制定了一种策略来通过在合适的细胞系中表达克隆受体并在添加组织提取物的分数后监测受体的激活来隔离这种天然配体。该策略最终将使我们能够净化活性分子并确定它们的结构。在此应用中，我们建议从与内皮素受体结构相关的两个这样的孤儿GPCR的天然配体分离，识别和表征药理学的天然配体。自然配体分离后，我们将通过克隆编码它们的前体蛋白并分析其在大脑和外围器官中的分布来研究其药理作用，研究其生物合成。这些受体没有结合已知的内皮蛋白肽，但在各种物种的进化过程中一直保持保守，表明它们必须发挥重要作用。这两种受体在大脑中都高度表达，其中一个受体最近被确定为帕金蛋白的底物，帕金（Parkin）是帕金森氏病的常染色体隐性形式突变的基因产物。天然配体的识别将使我们能够在正常和病理条件下研究这两个孤儿GPCR的生理功能。