Dynamic Mechanisms of Membrane Channel Gating by CryoEM

项目摘要 成孔膜通道是许多复杂生物现象的中心介质;例如 同步我们心脏的收缩和我们大脑中的电化学信号，检测光，声音， 触觉，味觉和嗅觉的变化这种能力依赖于用于 在空间和时间上调节其细胞活性。我们的研究小组致力于了解 这些类型的现象是由细胞间通信的非常复杂的策略编排的， 通过差距结。我们的目标是发展一个分子和原子水平的机械理解， 缝隙连接是如何协调细胞间通讯的为了达到这种细节水平，我们正在结合 电子冷冻显微镜（CryoEM）的独特功能，以及靶向生物物理和功能 研究来解决几个基本问题，如：i）差距结如何选择性地控制 细胞间的化学信息流ii）它们的活性如何被生理学上的变构调节 线索？iii）如何使用细胞信号平台来有效地控制天然多细胞环境中的通道活性。 环境？尽管它们具有生理和医学相关性，但膜蛋白仍然仅占~4%。 蛋白质结构数据库。然而，高分辨率单粒子领域的最新进展 CryoEM，加上膜蛋白生物化学的进步，开始彻底改变 我们对这些蛋白质的结构特征的描述。有了这些技术工具，我们正在解决 关于缝隙连接选择性和调节的几个关键问题。我们的调查结果是 期望提供开发所需的架构框架和机械知识 针对一系列缝隙连接相关疾病的靶向治疗，如失明，耳聋， 心律失常中风和癌症

Project Summary Pore-forming membrane channels are central mediators of many complex biological phenomena; such as synchronizing the contraction of our heart and electro-chemical signals in our brain, and detecting light, sound, touch, taste and smells of the world around us. This ability is dependent upon dynamic mechanism used to spatially and temporally modulate their cellular activity. Our research group is focused on understanding how these types of phenomena are choreographed by remarkably complex strategies of cell-to-cell communication, through the gap junctions. We aim to develop a molecular and atomic-level of mechanistic understanding of how gap junctions coordinate inter-cellular communication. To achieve this level of detail, we are combining the unique power of electron cryo-microscopy (CryoEM), together with targeted biophysical and functional studies to address several fundamental questions, such as: i) How do the gap junctions selectively control the flow of chemical information between cells? ii) How are their activities allosterically modulated by physiological cues? iii) How are cell-signaling platforms used to effectively control channel activity in a native multi-cellular environment? Despite their physiological and medical relevance, membrane proteins still only represent ~4% of the protein structure database. However, recent advances in the field of high-resolution single particle CryoEM, coupled with advancements in membrane protein biochemistry, are beginning to revolutionize the way we structurally characterize these proteins. With these technological tools in hand, we are addressing several key questions about gap junction selectivity and regulation. The results of our investigations are expected to provide an architectural framework and the mechanistic knowledge required for the development of targeted therapies against a range of gap junction related diseases, such as blindness, deafness, arrhythmia, stroke and cancers.!