Purification & Structural Analysis/Cannabinoid Receptor

描述：（申请人提供） 关于“大麻受体”的结构信息是有限的。这是 不幸的是，因为大麻是最广泛使用的街头毒品之一， 越来越多地被认为具有治疗作用。更多详细知识 关于这种受体的结构将大大有助于设计 激活和抑制药物，并促进治疗方案。 缺乏大麻结构信息的主要原因 受体（神经元大麻素受体，称为CB 1）是CB 1是 G蛋白偶联受体（GPCR）。GPCR蛋白是一个大家族， 膜受体蛋白质参与信号传递， 膜。不幸的是，这些蛋白质通常以低水平表达， 和大多数膜蛋白一样，很难纯化和操作。 部分得益于我们的经验净化和进行物理研究， 视紫红质（唯一的GPCR，其高分辨率晶体结构是 已知），我们提出表达和纯化大量的CB 1受体。 获得大量的纯CB 1将使我们能够尝试结晶CB 1， 也将使我们能够开始研究可能发生的动态变化 配体结合后的CB 1结构。这些研究将涉及使用 荧光和EPR光谱，以确定CB 1激活是否涉及 在其跨膜螺旋之一，螺旋F处的构象变化。这样的 先前已经在视紫红质和β-肾上腺素能神经元中观察到运动。 受体，如果在CB 1中检测到，将表明这些不同的GPCR可能共享 一种普遍的激活机制

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.