Electrostatic influence on IL-1 transport through the GSDMD pore

A variety of signals, including inflammasome activation, trigger the formation of large transmembrane pores by gasdermin D (GSDMD). There are primarily two functions of the GSDMD pore, to drive lytic cell death, known as pyroptosis, and to permit the release of leaderless interleukin-1 (IL-1) family cytokines, a process that does not require pyroptosis. We are interested in the mechanism by which the GSDMD pore channels IL-1 release from living cells. Recent studies revealed that electrostatic interaction, in addition to cargo size, plays a critical role in GSDMD-dependent protein release. Here, we determined computationally that to enable electrostatic filtering against pro-IL-1 beta, acidic lipids in the membrane need to effectively neutralize positive charges in the membrane-facing patches of the GSDMD pore. In addition, we predicted that salt has an attenuating effect on electrostatic filtering and then validated this prediction using a liposome leakage assay. A calibrated electrostatic screening factor is necessary to account for the experimental observations, suggesting that ion distribution within the pore may be different from the bulk solution. Our findings corroborate the electrostatic influence of IL-1 transport exerted by the GSDMD pore and reveal extrinsic factors, including lipid and salt, that affect the electrostatic environment.

Automated summary

In this study, computational simulation and experimental validation were used to investigate the mechanism by which the GSDMD pore channels IL-1 release from living cells. It was found that acidic lipids effectively neutralize positive charges in the membrane-facing patches of the GSDMD pore, enabling electrostatic filtering against pro-IL-1 beta. Additionally, it was discovered that salt has an attenuating effect on electrostatic filtering. These findings provide important insights into the electrostatic influence of IL-1 transport exerted by the GSDMD pore and the impact of extrinsic factors on the electrostatic environment.