Neuronal Gap Junctions: Cx36 Gating, Binding & Function

描述（由申请人提供）：神经元之间的电突触是 由间隙连接通道形成，离子和代谢物通过间隙连接通道 直接从一个单元格传递到下一个单元格。最近的证据表明，其他 蛋白质与间隙连接处的连接蛋白相关，我们将其称为 这种大分子复合物就是 Nexus。我们假设 Nexus 组件 可以调节间隙连接通道的特性，也可以 在细胞内信号转导中发挥作用。一种新的间隙连接蛋白， connexin36 已被鉴定其表达在很大程度上受到限制 神经元，我们已经证明由这种连接蛋白形成的通道具有 这些特性似乎独特地适合它作为神经元连接蛋白。的目标 该拨款申请旨在描述浇口、渗透性和 关于所涉及的蛋白质结构域的电导特性，以识别 大脑和神经元裂解物中与 connexin36 结合的其他蛋白质，测量 使用表面等离子体共振相互作用的强度，并执行 对神经元样细胞进行生理实验以确定 Cx36 的作用 神经元分化和基因表达模式的表达。在 此外，该应用程序的一个主要目标是获取结构信息 使用光谱方法分析与每个相互作用的 Cx36 域 其他以及与其他蛋白质。这些研究采用多学科方法 旨在探索该领域的新概念，因此预计 导致对间隙连接的调节和作用的新理解 神经系统。

DESCRIPTION (provided by applicant): Electrical synapses between neurons are formed by gap junction channels, through which ions and metabolites pass directly from one cell to the next. Recent evidence indicates that other proteins are associated with connexins at gap junctions, and we have termed this macromolecular complex the Nexus. We hypothesize that the Nexus components may regulate both the properties of the gap junction channels and also may function in intracellular signal transduction. A new gap junction protein, connexin36, has been identified that is largely restricted in its expression to neurons, and we have shown that the channels formed by this connexin have properties that seem to uniquely suit it as a neuronal connexin. The goals of this grant application are to characterize the gating, permeability and conductance properties with regard to the protein domains involved, to identify other proteins in brain and neuron lysates that bind to connexin36, measure the strengths of interaction using surface plasmon resonance, and perform physiological experiments on neuron-like cells to determine the effects of Cx36 expression on neuronal differentiation and gene expression patterns. In addition, a major goal of this application is to obtain structural information using spectroscopic methods regarding domains of Cx36 that interact with each other and with other proteins. These studies use a multidisciplinary approach directed at exploring a new concept in the field and as such are expected to lead to novel understanding of regulation and roles of gap junctions in the nervous system.