ATP and large signaling metabolites flux through caspase-activated Pannexin 1 channels

Pannexin 1 (Panx1) is a membrane channel implicated in numerous physiological and pathophysiological processes via its ability to support release of ATP and other cellular metabolites for local intercellular signaling. However, to date, there has been no direct demonstration of large molecule permeation via the Panx1 channel itself, and thus the permselectivity of Panx1 for different molecules remains unknown. To address this, we expressed, purified, and reconstituted Panx1 into proteoliposomes and demonstrated that channel activation by caspase cleavage yields a dye-permeable pore that favors flux of anionic, large-molecule permeants (up to similar to 1 kDa). Large cationic molecules can also permeate the channel, albeit at a much lower rate. We further show that Panx1 channels provide a molecular pathway for flux of ATP and other anionic (glutamate) and cationic signaling metabolites (spermidine). These results verify large molecule permeation directly through caspase-activated Panx1 channels that can support their many physiological roles.

Automated summary

The study demonstrated that Panx1 channels activated by caspase allow large molecule permeation, including anionic and large-molecule permeants, while also allowing large cationic molecules to permeate at a slower rate. Additionally, Panx1 channels provide a pathway for the flux of ATP and other signaling metabolites, both anionic (glutamate) and cationic (spermidine).