Synthetic inhibitors of the serine protease DPP9 activate the related NLRP1 and CARD8 inflammasomes and stimulate powerful innate immune responses. Thus, it seems plausible that a biomolecule similarly inhibits DPP9 and triggers inflammasome activation during infection, but one has not yet been discovered. Here, we wanted to identify and characterize DPP9-binding proteins to potentially uncover physiologically relevant mechanisms that control DPP9 s activity. Notably, we found that the redox sensor protein KEAP1 binds to DPP9 in an inactive conformation and stabilizes this non-native fold. At the same time, this inactive form of DPP9 reciprocally inhibits the ability of KEAP1 to bind to and degrade the transcri... More
Synthetic inhibitors of the serine protease DPP9 activate the related NLRP1 and CARD8 inflammasomes and stimulate powerful innate immune responses. Thus, it seems plausible that a biomolecule similarly inhibits DPP9 and triggers inflammasome activation during infection, but one has not yet been discovered. Here, we wanted to identify and characterize DPP9-binding proteins to potentially uncover physiologically relevant mechanisms that control DPP9 s activity. Notably, we found that the redox sensor protein KEAP1 binds to DPP9 in an inactive conformation and stabilizes this non-native fold. At the same time, this inactive form of DPP9 reciprocally inhibits the ability of KEAP1 to bind to and degrade the transcription factor NRF2, thereby inducing an antioxidant response. Although we discovered several experimental conditions, for example new protein expression and chemical denaturation, that force DPP9 out of its folded dimeric state and into a KEAP1-binding state, the key danger-related stimulus that causes this critical DPP9 conformational change is not yet known. Regardless, our data now reveal that an endogenous DPP9 inhibition mechanism does in fact exist, and moreover that DPP9, like the other NLRP1 regulator thioredoxin-1, is directly coupled to the intracellular redox potential. Overall, we expect this work will provide the foundation to discover additional biomolecules that regulate DPP9 s activity, the DPP9-KEAP1 interaction, the intracellular redox environment, and the NLRP1 and CARD8 inflammasomes.
