Neuronal Gap Junctions: Cx36 Gating, Binding & Function

描述(申请人提供)：神经元之间的电突触是 由缝隙连接通道形成，离子和代谢物通过该通道 直接从一个细胞传到下一个细胞。最近的证据表明，其他 蛋白质与缝隙连接蛋白有关，我们称之为 这个大分子复合体就是Nexus。我们假设Nexus组件 可以调节间隙结沟道的属性，并且还可以 在细胞内信号转导中的作用。一种新的缝隙连接蛋白， 连接蛋白36已被发现，其表达在很大程度上受限于 神经元，我们已经证明了由这种连接蛋白形成的通道 作为一种神经元连接蛋白似乎特别适合它的特性。的目标 这项拨款申请是为了表征浇注、渗透性和 与所涉及的蛋白质结构域有关的电导特性，以识别 大脑和神经元裂解物中与连接蛋白36结合的其他蛋白质，测量 使用表面等离子激元共振的相互作用强度，并执行 神经元样细胞的生理实验确定Cx36的作用 神经元分化的表达和基因表达模式。在……里面 此外，此应用程序的一个主要目标是获取结构信息 使用关于与每个相互作用的Cx36结构域的光谱方法 其他以及与其他蛋白质的结合。这些研究使用了多学科方法。 旨在探索该领域的新概念，并因此有望 导致对缝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.