Structural Studies on the Cannabinoid Receptor

大麻素受体的结构研究

• 批准号: 6640565
• 负责人: STEVEN E MANSOOR
• 金额: $ 2.95万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-08-01 至 2007-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6640565
• 关键词: G protein; SDS polyacrylamide gel electrophoresis; affinity labeling; animal tissue; autoradiography; behavioral /social science research tag; beta adrenergic receptor; cannabinoid receptor; chimeric proteins; conformation; electron spin resonance spectroscopy; fluorescence spectrometry; fluorescent dye /probe; inhibitor /antagonist; marijuana abuse; nucleotides; polymerase chain reaction; predoctoral investigator; protein binding; protein structure function; receptor binding; receptor coupling; receptor expression; stimulant /agonist

DESCRIPTION (provided by applicant): The long-term goal of the proposed research is to understand mechanistically how the neuronal cannabinoid receptor (CB1) becomes activated. Several aspects of this question will be explored. Below is a brief description of my specific aims. (Aim 1) The most widely accepted model for G-protein coupled receptor (GPCR) activation is the two-state model yet recent evidence suggests that this model is too simple to describe GPCR activation. The validity of the two-state model for CB1 activation will be assessed using fluorescence spectroscopy to monitor conformational states of the activated receptor to determine if there is one active state conformation or multiple, distinct conformational states. (Aim 2) Nucleotide binding to docked G-proteins has been shown to affect a GPCR's affinity for its ligands. By exploring whether this is true for the CB1 receptor, I will determine if nucleotide allosteric regulation of ligand binding works through inducing conformational changes in the receptor. (3) The mechanisms by which CB1 is constitutively active will be examined by attempting to make mutations in the receptor that reduce or abolish the constitutive activity. These mutants will then be studied structurally to examine if and how any conformational changes are linked to constitutive activity. Because marijuana is a widely used street drug, and the cannabinoid receptor is a member of the G-protein coupled receptor family (the largest class of drug discovery targets), understanding the mechanism by which the CB1 receptor is modulated is important both clinically and scientifically.

描述（由申请人提供）：拟议的研究的长期目标是从机械上理解神经元大麻素受体（CB1）如何被激活。 将探讨这个问题的几个方面。 以下是对我的具体目标的简要描述。 （AIM 1）G蛋白偶联受体（GPCR）激活最广泛接受的模型是两态模型，但最近的证据表明，该模型太简单了，无法描述GPCR激活。 两态模型对CB1激活的有效性将使用荧光光谱进行评估，以监测活化受体的构象状态，以确定是否存在一个活性状态构象或多个不同的构象状态。 （AIM 2）已证明与对接G蛋白的核苷酸结合会影响GPCR对其配体的亲和力。 通过探索CB1受体是否正确，我将通过诱导受体构象变化来确定配体结合的核苷酸变构调节。 （3）将通过尝试在受体中减少或废除本构活性的受体突变来检查CB1具有组成性活性的机制。 然后将在结构上研究这些突变体，以检查是否以及如何与本构活性相关联。由于大麻是一种广泛使用的街道药物，并且大麻素受体是G蛋白偶联受体家族（最大的药物发现靶标类）的成员，因此了解CB1受体被调节的机制在临床上和科学上都很重要。