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Functioning of Cx43 Hemichannels Demonstrated by Single Channel Properties

机译:单通道属性展示的Cx43半通道的功能

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It has been suggested that the opening of non-junctional connexin 43 (Cx43) hemichannels may play a role in cell physiology, but some workers doubt the reality of hemichannel openings. Mere we show data on unitary conductance and voltage gating properties demonstrating that Cx43 hemichannels can open. Membrane depolarization > +60 mV induced single hemichannel currents in HeLa cells expressing Cx43 or Cx43 with enhanced green fluorescent protein attached to the carboxy terminal (Cx43-EGFP). The conductance of single hemichannels was ~220 pS, about twice that of the cell-cell channels. Cx43 and Cx43-EGFP hemichannels exhibited slow transitions (>5 ms) between closed and fully open states. Cx43 hemichannels also exhibited fast transitions (< 1 ms) between the fully open state and a substate of ~75 pS. Similar gating was described for their respective cell-cell channels. No comparable single channel activity was detected in the parental (nontransfected cells) or HeLa cells expressing Cx43 fused at the amino terminal with EGFP (EGFP-Cx43). The latter chimera was inserted into the surface and formed plaques, but did not express functional hemichannels or cell-cell channels. These data convincingly demonstrate the opening of Cx43 hemichannels.
机译:有人提出非连接连接蛋白43(Cx43)半通道的开放可能在细胞生理学中发挥作用,但一些工人怀疑半通道开放的现实。我们仅显示有关单位电导和电压门控特性的数据,表明Cx43半通道可以打开。膜去极化> +60 mV会在表达Cx43或Cx43的HeLa细胞中诱导单个半通道电流,而Cx43或Cx43的绿色荧光蛋白附着在羧基末端(Cx43-EGFP)。单个半通道的电导约为220 pS,约为细胞通道的两倍。 Cx43和Cx43-EGFP半通道在关闭状态和完全打开状态之间显示出缓慢的跃迁(> 5 ms)。 Cx43半通道在完全打开状态和〜75 pS的子状态之间也表现出快速的跃迁(<1 ms)。对于它们各自的细胞通道描述了类似的门控。在表达在氨基末端与EGFP融合的Cx43的亲本(未转染的细胞)或HeLa细胞中未检测到可比的单通道活性(EGFP-Cx43)。将后者的嵌合体插入表面并形成噬斑,但是不表达功能性半通道或细胞-细胞通道。这些数据令人信服地证明了Cx43半通道的开放。

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