Purification & Structural Analysis/Cannabinoid Receptor

描述：(申请人提供) 关于“大麻受体”的结构信息有限。这是 不幸的是，大麻是最广泛使用的街头毒品之一，而且 越来越多的人将其归因于治疗属性。更详细的知识 关于这种受体的结构将极大地帮助两者的设计 激活和抑制药物，促进治疗制度。 缺乏关于大麻的结构性信息的一个主要原因 受体(神经性大麻素受体，称为CB1是指CB1是 G蛋白偶联受体(GPCR)。GPCR蛋白是一大家族 参与信号传递的膜受体蛋白 膜。不幸的是，这些蛋白质通常在低水平表达， 和大多数膜蛋白一样，它们很难纯化和操作。 部分得益于我们净化和开展物理研究的经验 视紫红质(唯一具有高分辨率晶体结构的GPCR 已知)我们建议表达和纯化大量的CB1受体。 获得大量的纯CB1将允许我们尝试结晶CB1， 还将使我们能够开始研究可能发生在 配体结合时的CB1结构。这些后一项研究将涉及使用 荧光和EPR光谱来确定CB1的激活是否涉及 它的一个跨膜螺旋螺旋F的构象变化。 以前在视紫红质和β-肾上腺素能神经元中观察到运动 受体，如果在CB1中检测到，将表明这些不同的GPCR可能共享 一种通用的激活机制。

DESCRIPTION: (provided by the applicant) Structural information about the "marijuana receptor" is limited. This is unfortunate since marijuana is one of the most widely used street drugs, and is increasingly being ascribed therapeutic properties. More detailed knowledge about the structure of this receptor would greatly assist the design of both activating and inhibiting drugs, and facilitate treatment regimes. A major reason for the lack of structural information about the marijuana receptor (the neuronal cannabinoid receptor, called CB1 is the fact that CB1 is a G-protein coupled receptor (GPCR). The GPCR proteins are a large family of membrane receptor proteins that are involved in transmitting signals across membranes. Unfortunately, these proteins are usually expressed at low levels, and like most membrane proteins, are difficult to purify and manipulate. Aided in part by our experience purifying and carrying out physical studies of rhodopsin (the only GPCR for which a high resolution crystal structure is known) we propose to express and purify large amounts of the CB1 receptor. Access to large quantities of pure CB1 will allow us to try crystallizing CB1, and will also enable us to begin studying dynamic changes that may occur in the CB1 structure upon ligand binding. These latter studies will involve using fluorescence and EPR spectroscopy to determine if CB1 activation involves a conformational change at one of its transmembrane helices, helix F. Such a movement has previously been observed in rhodopsin and the beta-adrenergic receptor, and if detected in CB1, will suggest these disparate GPCRs may share a universal mechanism of activation.