Purified Serotonin Receptors for Structural Studies

用于结构研究的纯化血清素受体

• 批准号: 6690933
• 负责人: David Salom
• 金额: $ 14.89万
• 依托单位: NOVASITE PHARMACEUTICALS, INC.
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-08-01 至 2004-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6690933
• 关键词: Xenopus; Xenopus oocyte; cell surface receptors; method development; protein purification; protein structure; rod cell; serotonin receptor; three dimensional imaging /topography

DESCRIPTION (provided by applicant): Serotonin (5-HT) is a neurotransmitter that has been implicated in the aetiology of numerous Mental Health disorders, including depression, anxiety, social phobia, schizophrenia, obsessive-compulsive and panic disorders, migraine, and eating disorders. There are 12 5-HT receptor subtypes with similar recognition properties yet widely divergent physiological roles that belong to the same family of G Protein Coupled Receptors (GPCRs), hindering the discovery of subtype-selective drugs. Knowledge of the 3-dimensional structure of these receptors would enable the design of selective drugs. However, solving the structure at atomic resolution of membrane proteins such as GPCRs has been hampered by the difficulty to purify sufficient amounts of homogeneous and functional protein. Here, it is proposed a novel, proprietary expression system combined with a purification method to generate large amounts of high quality purified receptor, which could solve the bottleneck for structural elucidation of these important drug targets. In Phase I, the feasibility of the proposed approach will be tested for all the 12 human 5-HT GPCRs. In Phase II, this approach will be developed resulting in purified homogenous protein for each human 5- HT receptor subtype. This high quality purified material will be a highly valuable product enabling structural elucidation of these receptors.

描述（由申请人提供）：5-羟色胺（5-HT）是一种神经递质，与许多精神健康障碍的病因有关，包括抑郁症、焦虑症、社交恐惧症、精神分裂症、强迫症和恐慌症、偏头痛和进食障碍。有12种5-HT受体亚型具有相似的识别特性，但生理作用差异很大，属于同一家族的G蛋白偶联受体（GPCR），阻碍了亚型选择性药物的发现。了解这些受体的三维结构将使选择性药物的设计成为可能。然而，以原子分辨率解析膜蛋白如GPCR的结构受到难以纯化足够量的均质和功能性蛋白质的阻碍。在这里，它提出了一种新的，专有的表达系统与纯化方法相结合，以产生大量的高质量的纯化受体，这可以解决这些重要的药物靶点的结构解析的瓶颈。在第一阶段，将对所有12个人5-HT GPCR测试所提出的方法的可行性。在II期，将开发这种方法，从而产生用于每个人5- HT受体亚型的纯化的同质蛋白。这种高质量的纯化材料将是一个非常有价值的产品，使这些受体的结构说明。

项目成果

Expression of functional G protein-coupled receptors in photoreceptors of transgenic Xenopus laevis.

转基因非洲爪蟾光感受器中功能性 G 蛋白偶联受体的表达。

• DOI: 10.1021/bi051386z
• 发表时间: 2005
• 期刊: Biochemistry.
• 作者: Zhang,Li;Salom,David;He,Jianhua;Okun,Alex;Ballesteros,Juan;Palczewski,Krzysztof;Li,Ning
• 通讯作者: Li,Ning

Heterologous expression and purification of the serotonin type 4 receptor from transgenic mouse retina.

转基因小鼠视网膜中 4 型血清素受体的异源表达和纯化。

• DOI: 10.1021/bi8018527
• 发表时间: 2008
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: Salom,David;Wu,Nan;Sun,Wenyu;Dong,Zhiqian;Palczewski,Krzysztof;Jordan,Steven;Salon,JohnA
• 通讯作者: Salon,JohnA