Abstract:
Connexins are the proteins that form the gap junction channels that are essential for cell-to-cell communication. These channels are formed by head-to-head docking of hemichannels (each from one of two adjacent cells). Free \"undocked\" hemichannels at the plasma membrane are mostly closed, although they are still important under physiological conditions. However, abnormal and sustained increase in hemichannel activity due to connexin mutations or acquired conditions can produce or contribute to cell damage. For example, mutations of Cx26, a connexin isoform, can increase hemichannel activity and cause deafness. Studies using purified isolated systems under well-controlled conditions are essential for a full understanding of molecular mechanisms of hemichannel function under normal conditions and in disease, and here, we present methodology for the expression, purification, and functional analysis of hemichannels formed by Cx26.
摘要:
连接蛋白是形成间隙连接通道的蛋白质,对于细胞间通讯至关重要。这些通道通过半通道(每个来自两个相邻单元中的一个)的头对头对接形成。质膜上的自由“未对接”半通道大多是封闭的,尽管它们在生理条件下仍然很重要。然而,由于连接蛋白突变或获得性条件引起的半通道活性的异常和持续增加可以产生或有助于细胞损伤。例如,Cx26的突变,一种连接蛋白亚型,会增加半通道活动并导致耳聋。在良好控制的条件下使用纯化的分离系统进行的研究对于充分了解正常条件下和疾病中半通道功能的分子机制至关重要。在这里,我们提出了表达式的方法,净化,和Cx26形成的半通道的功能分析。