STRUCTURE/FUNCTION RELATIONSHIPS OF THE CCK B RECEPTOR

CCK B 受体的结构/功能关系

• 批准号: 2146014
• 负责人: ALAN S KOPIN
• 金额: $ 19.67万
• 依托单位: TUFTS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-09-30 至 1997-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2146014
• 关键词: G protein; affinity labeling; brain cell; chimeric proteins; cholecystokinin; complementary DNA; gene deletion mutation; human genetic material tag; human tissue; laboratory rabbit; molecular cloning; neoplastic cell culture for noncancer research; neuropeptide receptor; neuropharmacology; nucleic acid hybridization; oligonucleotides; polymerase chain reaction; protein structure function; receptor binding; receptor coupling; receptor expression; second messengers; site directed mutagenesis

The cholecystokinin (CCK) type B receptor in brain plays an important role in modulating anxiety and panic attacks. CCK-B receptor antagonists have been shown to block the neuronal response to CCK and have been proposed as a potentially efficacious new class of anti-anxiety medication. Isolation of a cloned CCK-B receptor and expression in a heterologous cell line, would expedite the development of this new class of drugs. We have isolated a cDNA encoding the first cloned member of the gastrin/CCK receptor family and have evidence that our receptor is highly homologous to the brain CCK-B receptor. Understanding the link between structure and function of the CCK-B receptor will allow more rational design of receptor antagonists. Determination of whether the "CCK-B receptor" (like the biogenic amine receptors) represents a family of related receptors, each with characteristic pharmacology, rather than a single receptor, would allow development of drugs targeted to a specific receptor subtype in brain. We have brought together a consortium of individuals with expertise in molecular biology, pharmacology, protein chemistry, and receptor characterization to undertake the isolation, expression, and characterization of the CCK-B and other related brain receptors. Collectively this group is experienced in all aspects of the proposed project and have been working together for the past year on the cloning and characterization of the gastrin receptor (PNAS, 1992, in press). The first objective is to isolate the CCK-B and related receptors from brain by low stringency hybridization and PCR with degenerate oligonucleotides. The CCK-B and related receptors will ten be pharmacologically characterized to determine ligand binding specificity and the second messenger pathways which couple to these receptors. The ligand binding sites will be mapped with photoaffinity labels which we have previously used in characterizing other members of this receptor family as well as with a series of new intrinsic photoaffinity probes which we will develop specifically for the CCK-B receptor.

大脑中B型B受体的胆囊动蛋白（CCK）起重要作用 在调节焦虑和恐慌发作中。 CCK-B受体拮抗剂具有 已显示可阻止神经元对CCK的反应，并已被提出为 一种潜在有效的新型抗焦虑药。 隔离 克隆的CCK-B受体和异源细胞系中的表达， 将加快这种新毒品的发展。 我们已经隔离了编码胃蛋白/CCK的第一个克隆成员的cDNA 受体家族并有证据表明我们的受体高度同源 脑CCK-B受体。 了解结构和 CCK-B受体的功能将允许更合理的受体设计 对手。 确定是否“ CCK-B受体”（例如 生物胺受体）代表一个相关受体家族，每个受体家族 具有特征性药理学，而不是单个受体，将 允许开发针对特定受体亚型的药物 脑。 我们将一个具有专业知识的个人联盟汇集在一起 分子生物学，药理学，蛋白质化学和受体 表征要进行隔离，表达和 CCK-B和其他相关大脑受体的表征。 该小组在提议的各个方面都有经验 项目并在过去的一年中一直在克隆和 胃蛋白受体的表征（PNAS，1992，印刷中）。 第一个目的是将CCK-B和相关受体与 大脑通过低严格的杂交和退化的PCR 寡核苷酸。 CCK-B和相关受体将十个 在药理特征以确定配体结合特异性和 夫妇与这些受体的第二个信使途径。 配体 绑定站点将用我们拥有的光附属物标签映射 以前用于表征该受体家族的其他成员 以及一系列新的固有的光性探针，我们将 专为CCK-B受体开发。