STRUCTURE-FUNCTION RELATIONSHIPS OF THE CCK-B RECEPTOR

CCK-B 受体的结构-功能关系

• 批准号: 3248116
• 负责人: ALAN S KOPIN
• 金额: $ 13.88万
• 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/3248116
• 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.

脑内的胆囊收缩素（CCK）B型受体在胆囊收缩的发生、发展中起着重要的作用 调节焦虑和恐慌发作 CCK-B受体拮抗剂具有 已显示阻断神经元对CCK的反应，并已被提议作为 一种可能有效的新型抗焦虑药物 隔离 克隆的CCK-B受体并在异源细胞系中表达， 会加速这类新药的研发 我们已经分离了编码第一个克隆的胃泌素/CCK成员的cDNA， 受体家族，并有证据表明我们的受体与 大脑CCK-B受体。 理解结构与 CCK-B受体的功能将允许更合理的受体设计 对手。 确定"CCK-B受体"（如 生物胺受体）代表一个相关受体家族，每个 具有特征药理学，而不是单一受体， 允许开发针对特定受体亚型的药物， 个脑袋 我们召集了一个由在以下方面具有专长的个人组成的财团： 分子生物学、药理学、蛋白质化学和受体 表征进行隔离，表达，和 CCK-B和其他相关脑受体的表征。 总的来说，该团队在提议的各个方面都经验丰富 项目，并一直在共同努力，在过去的一年里，克隆和 胃泌素受体的特征（PNAS，1992，出版中）。 第一个目的是分离CCK-B和相关受体， 通过低严格性杂交和简并 寡核苷酸 CCK-B和相关受体将在 表征以确定配体结合特异性的试剂盒， 与这些受体结合的第二信使途径。 配体 结合位点将被绘制成具有光亲和性的标记， 以前用于表征该受体家族的其他成员， 以及一系列新的内在光亲和探针，我们将 专门针对CCK-B受体开发。