School of Energy, Environmental, Biological and Medical Engineering, Engineering Research Center, University of Cincinnati, Cincinnati, Ohio, United States of America.
PLoS One. 2012;7(10):e47775. doi: 10.1371/journal.pone.0047775. Epub 2012 Oct 24.
Connexin43 (Cx43) is the most ubiquitous gap junction protein in the human body and is essential for cell-to-cell communication in a variety of organs and organ systems. As a result, Cx43 is responsible for mediating both electrical and chemical signals, passing dissolved solutes and small signaling molecules between cells in a coordinated fashion. Here, we explore the electrophysiological properties of hemichannels formed from Cx43 and Cx43 fused to eGFP (Cx43eGFP) and their interactions in a planar lipid membrane (BLM). Unlike in vivo patch clamp experiments, Cx43 was purified and isolated from other membrane constituents allowing elucidation of individual protein responses to various electrical and chemical stimuli. Using this system, we examined hemichannel electrophysiology and the roles of several well-known gap junction blockers, namely: lanthanum, heptanol, carbenoxalone and lindane. We also observed a critical number of hemichannels required for an accelerated conductance increase, an emergent electrical signature indicative of plaque formation.
间隙连接蛋白 43(Cx43)是人体内最普遍存在的缝隙连接蛋白,对于各种器官和系统中的细胞间通讯至关重要。因此,Cx43 负责介导电信号和化学信号,协调地在细胞之间传递溶解的溶质和小信号分子。在这里,我们探索了由 Cx43 和融合了 eGFP 的 Cx43(Cx43eGFP)形成的半通道的电生理特性及其在平面脂质膜(BLM)中的相互作用。与体内膜片钳实验不同,Cx43 是从其他膜成分中纯化和分离出来的,这使得我们能够阐明单个蛋白对各种电和化学刺激的反应。使用该系统,我们研究了半通道的电生理学特性以及几种众所周知的缝隙连接阻滞剂的作用,即:镧、庚醇、卡波酮和林丹。我们还观察到形成斑块的加速电导增加的半通道的临界数量,这是一种新兴的电特征。