Journal
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
Volume 24, Issue 14, Pages -Publisher
MDPI
DOI: 10.3390/ijms241411612
Keywords
Cx43; Panx1; channel gating; single-channel analysis
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Cx43 hemichannels (HCs) and Panx1 channels, two genetically distant protein families, have been debated for their similarities in expression, membrane topology, and electrical properties. This study used mimetic peptides Gap19 and (10)Panx1 to investigate their cross-effects on Cx43 HCs and Panx1 channels. The results showed that Gap19 significantly inhibited Panx1 channels, while high concentrations of (10)Panx1 significantly reduced the activity of Cx43 HCs.
Cx43 hemichannels (HCs) and Panx1 channels are two genetically distant protein families. Despite the lack of sequence homology, Cx43 and Panx1 channels have been the subject of debate due to their overlapping expression and the fact that both channels present similarities in terms of their membrane topology and electrical properties. Using the mimetic peptides Gap19 and (10)Panx1, this study aimed to investigate the cross-effects of these peptides on Cx43 HCs and Panx1 channels. The single-channel current activity from stably expressing HeLa-Cx43 and C6-Panx1 cells was recorded using patch-clamp experiments in whole-cell voltage-clamp mode, demonstrating 214 pS and 68 pS average unitary conductances for the respective channels. Gap19 was applied intracellularly while (10)Panx1 was applied extracellularly at different concentrations (100, 200 and 500 & mu;M) and the average nominal open probability (NPo) was determined for each testing condition. A concentration of 100 & mu;M Gap19 more than halved the NPo of Cx43 HCs, while 200 & mu;M (10)Panx1 was necessary to obtain a half-maximal NPo reduction in the Panx1 channels. Gap19 started to significantly inhibit the Panx1 channels at 500 & mu;M, reducing the NPo by 26% while reducing the NPo of the Cx43 HCs by 84%. In contrast (10)Panx1 significantly reduced the NPo of the Cx43 HCs by 37% at 100 & mu;M and by 83% at 200 & mu;M, a concentration that caused the half-maximal inhibition of the Panx1 channels. These results demonstrate that (10)Panx1 inhibits Cx43 HCs over the 100-500 & mu;M concentration range while 500 & mu;M intracellular Gap19 is necessary to observe some inhibition of Panx1 channels.
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