Cx43 Hemichannel and Panx1 Channel Modulation by Gap19 and ¹⁰Panx1 Peptides

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 ¹⁰Panx1, 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 ¹⁰Panx1 was applied extracellularly at different concentrations (100, 200 and 500 μM) and the average nominal open probability (NP_o) was determined for each testing condition. A concentration of 100 µM Gap19 more than halved the NP_o of Cx43 HCs, while 200 µM ¹⁰Panx1 was necessary to obtain a half-maximal NP_o reduction in the Panx1 channels. Gap19 started to significantly inhibit the Panx1 channels at 500 µM, reducing the NP_o by 26% while reducing the NP_o of the Cx43 HCs by 84%. In contrast ¹⁰Panx1 significantly reduced the NP_o of the Cx43 HCs by 37% at 100 µM and by 83% at 200 µM, a concentration that caused the half-maximal inhibition of the Panx1 channels. These results demonstrate that ¹⁰Panx1 inhibits Cx43 HCs over the 100-500 µM concentration range while 500 µM intracellular Gap19 is necessary to observe some inhibition of Panx1 channels.