Towards a structural and temporal understanding of phototransduction

G蛋白偶联受体（GPCR）介导的信号系统的机制仍然是 尽管这些系统在过去世纪受到了高度关注，但这些问题仍未得到解决。部分原因 在研究膜蛋白在其自然环境中的困难，通过提供分子生物学的方法， 续费目前建议的重点是应用新的生物物理方法来解开 GPCR介导的信号通路的分子和时间奥秘。视紫红质与视觉系统 这将是该研究计划的最初重点。这种原型GPCR信号传导系统提供了几种 这些优点将允许新的生物物理方法的应用。原子力显微镜将 产生单个分子的高分辨率图像， 系统的信息。单分子力谱将提供详细的信息， 视紫红质中稳定蛋白质并促进其功能的分子相互作用。低温电子 层析成像将进入未扰动的棒外节段，以提供关于这一点的结构信息。 在这个场所，它们的功能是通过细胞间和大分子来实现的。荧光共振 能量转移将用于检测信号蛋白的蛋白质-蛋白质相互作用，以监测信号蛋白的表达。 动态交互，定义了信号过程和发生信号过程的时间框架。一起 通过这种独特的方法学组合获得的信息将提供分子生物学的关键片段， 这些信息目前不适用于这些系统。这将有助于确定分子机制 潜在的信号事件，支配由GPCR调控的重要生理过程。 G蛋白偶联受体（GPCRs）代表了最大类的细胞表面蛋白和药物靶点 目前在市场上。这个蛋白质家族几乎参与了所有的生理过程， 这些系统的功能障碍会导致失明、成瘾、糖尿病和心脏病等疾病。 疾病尽管GPCR的重要性，其作用的准确分子描述仍然缺乏。 了解这些系统的分子奥秘将有助于开发更有效的 治疗解决方案。

The mechanism underlying G protein-coupled receptor (GPCR)-mediated signaling systems is still unresolved despite the intense focus these systems have received over the past century. This is due in part to the difficulties in studying membrane proteins in their native context by methods that provide molecular details. The focus of the current proposal is to apply novel biophysical methodologies to unravel the molecular and temporal mysteries of GPCR-mediated signaling pathways. Rhodopsin and the visual system will be the initial focus of the research program. This prototypical GPCR signaling system offers several advantages that will allow for the application of novel biophysical approaches. Atomic force microscopy will result in high-resolution images of individual molecules that will provide structural and organizational information of the system. Single-molecule force spectroscopy will provide detailed information on the molecular interactions in rhodopsin that stabilize the protein and promote its function. Cryo-electron tomography will gain access to an unperturbed rod outer segment to provide structural information on this compartment and on the macromolecules that carry out their function at this venue. Fluorescence resonance energy transfer will be utilized to detect protein-protein interactions of signaling proteins to monitor the dynamic interactions that define the signaling process and the timeframe in which this takes place. Together the information obtained by this unique combination of methodologies will provide key pieces of molecular information that is currently unavailable for these systems. This will help define the molecular mechanism underlying the signaling events that govern important physiological processes regulated by GPCRs. G protein-coupled receptors (GPCRs) represent the largest class of cell surface proteins and drug targets currently on the market. This family of proteins is involved in virtually every physiological process, and dysfunctions in these systems can lead to diseases such as blindness, addiction, diabetes, and heart disease. Despite the importance of GPCRs an accurate molecular description of their action is still lacking. Understanding the molecular mysteries of these systems will lead to the development of more effective therapeutic solutions.