Structural Dynamics of Retinal Binding and Release

视网膜结合和释放的结构动力学

• 批准号: 7094863
• 负责人: David L Farrens
• 金额: $ 26.85万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-06-01 至 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7094863
• 关键词: arrestins; rhodopsin

DESCRIPTION (provided by applicant): The long-term goal of our research is to understand the molecular mechanisms through which G-protein coupled receptors (GPCRs) are activated and attenuated. These receptors represent the largest family in the human genome, and most importantly, are the target of most pharmaceutical drugs. Our studies focus primarily on the GPCR rhodopsin and its affiliate proteins. Although knowledge of the proteins involved in visual signaling has been enormously advanced through recent crystallographic studies, the critical structural changes they undergo during activation and attenuation remain largely a matter of speculation. In particular, we lack even the most rudimentary information about the dynamic events that occur during the attenuation of rhodopsin signaling, namely, the mechanisms used to release retinal from the opsin-binding pocket, and how these processes affect arrestin binding and release. Understanding these processes is of fundamental importance for vision research, as the stability of the retinal linkage varies widely among different opsins and is a factor in some visual disease states. Furthermore, little is known regarding the mechanism and kinetics of arrestin release from activated rhodopsin. In Aim I of this proposal we will determine the mechanisms through which rhodopsin controls the hydrolysis of its retinal Schiff base linkage. In Aim II we will use our expertise in site-directed labeling methods to examine dynamic and structural changes occurring in the extracellular loop region of rhodopsin as it binds and releases retinal. Finally, in Aim III, we will utilize a new assay we have developed to study arrestin release from activated rhodopsin. Understanding what makes arrestin "let go" after binding rhodopsin is fundamentally important, since stable rhodopsin-arrestin complexes have been suggested to be a contributing factor in apoptosis and autosomal dominant retinitis pigmentosa (ADRP). Furthermore, using our new assay, we find intriguing evidence that arrestin binds to and traps a post-Meta II photodecay product, possibly Meta III. Thus, arrestin may also serve to limit the release of free retinal under bright light conditions, and thus help limit the formation of oxidative retinal adducts that can contribute to diseases like atrophic age-related macular degeneration (AMD).

描述（由申请人提供）：我们研究的长期目标是了解 G 蛋白偶联受体 (GPCR) 被激活和减弱的分子机制。这些受体代表了人类基因组中最大的家族，最重要的是，它们是大多数药物的靶标。我们的研究主要集中在 GPCR 视紫红质及其附属蛋白。尽管通过最近的晶体学研究，对参与视觉信号传导的蛋白质的了解已经取得了巨大进展，但它们在激活和减弱过程中经历的关键结构变化在很大程度上仍然是猜测的问题。特别是，我们甚至缺乏关于视紫红质信号传导减弱期间发生的动态事件的最基本信息，即用于从视蛋白结合袋释放视网膜的机制，以及这些过程如何影响视紫红质抑制蛋白的结合和释放。了解这些过程对于视觉研究至关重要，因为不同视蛋白之间视网膜连接的稳定性差异很大，并且是某些视觉疾病状态的一个因素。此外，关于从激活的视紫红质释放视紫红质抑制蛋白的机制和动力学知之甚少。在本提案的目标一中，我们将确定视紫红质控制其视网膜希夫碱键水解的机制。在目标 II 中，我们将利用我们在定点标记方法方面的专业知识来检查视紫红质结合和释放视网膜时细胞外环区域发生的动态和结构变化。最后，在目标 III 中，我们将利用我们开发的新测定法来研究激活视紫红质中抑制蛋白的释放。了解抑制蛋白在结合视紫红质后“释放”的原因至关重要，因为稳定的视紫红质-抑制蛋白复合物已被认为是细胞凋亡和常染色体显性视网膜色素变性（ADRP）的影响因素。此外，使用我们的新检测方法，我们发现了有趣的证据，证明视紫红质抑制蛋白结合并捕获 Meta II 后的光衰产物，可能是 Meta III。因此，视紫红质抑制蛋白还可能有助于限制亮光条件下游离视网膜的释放，从而有助于限制氧化性视网膜加合物的形成，氧化性视网膜加合物可能导致萎缩性年龄相关性黄斑变性（AMD）等疾病。