Structural basis for ADAM17 activation by the iRhom1 pseudoprotease

The a disintegrin and metalloproteinase (ADAM)-17 releases pro-inflammatory cytokines, including tumor necrosis factor-α, and several epidermal growth factor receptor ligands from cells. ADAM17 is post-translationally regulated by the inactive rhomboid pseudoproteases iRhom1 and iRhom2, and dysregulation of this signaling axis contributes to diseases ranging from chronic inflammation to cancer. Here, we present the cryo-electron microscopy structure of the ADAM17 zymogen bound to iRhom1, revealing structural features essential for complex formation and protease activation. We identify a transmembrane α-helix and conserved cytoplasmic element in iRhom1, termed the re-entry loop, that functions as a molecular relay transmitting intracellular signals across the membrane to activate ADAM17. We also demonstrate that a human disease-associated iRhom1 mutation linked to cardiomyopathy disrupts ADAM17 maturation and trafficking. Finally, we apply all-atom molecular dynamics simulations to model the mature and active ADAM17-iRhom1 complex. These findings provide mechanistic insights into how iRhom1 regulates ADAM17 function across the membrane.