The chapter discusses the role of post-translational modifications (PTMs) in regulating NLRP3 inflammasome signaling. PTMs, including phosphorylation, ubiquitination, SUMOylation, acetylation, and glycosylation, influence the activity of inflammasome components through effects on protein-protein interactions, stability, and localization. The review highlights how these modifications can both activate and inhibit inflammasome signaling, contributing to the regulation of inflammatory responses. Key points include: 1. **Non-transcriptional Priming**: Inflammasome activation requires two signals: a priming signal that drives NLRP3 expression and an activating signal that triggers its activation. PTMs like deubiquitination and phosphorylation play crucial roles in this process. 2. **NLRP3 Phosphorylation**: Phosphorylation at specific sites on NLRP3, such as Serine-5 and Serine-198, can inhibit or activate inflammasome activity, respectively. Kinases like AKT, JNK, and BTK are involved in these modifications. 3. **Ubiquitination and SUMOylation**: Ubiquitination at Lys 496 and SUMOylation at Lys 689 can promote or inhibit inflammasome activation, respectively. E3 ligases like TRIM31 and Cbl-b are involved in these modifications. 4. **Palmitoylation**: Palmitoylation can affect NLRP3 stability and protein-protein interactions, with zDHHC12 promoting degradation and zDHHC5 enhancing binding to NEK7. 5. **Other Modifications**: Acetylation, nitrosylation, and ADP-ribosylation also influence inflammasome activity, with specific modifications affecting cytokine release and cell death. 6. **Adaptor Protein ASC**: PTMs on ASC, such as phosphorylation and ubiquitination, regulate speck formation and inflammasome activation. 7. **Caspase-1**: Caspase-1 is modified by phosphorylation, ubiquitination, and nitrosylation, each influencing its activity and downstream signaling. 8. **Gasdermin D**: Gasdermin D is modified by phosphorylation, succination, palmitoylation, and oxidation, each contributing to its pore-forming and pyroptotic functions. 9. **IL-1β**: Pro-IL-1β is modified by glutathionylation and ubiquitination, affecting its stability and activity. The review concludes by highlighting outstanding questions about the integration of PTMs, the dominance of specific modifications, and the cooperative signaling pathways that regulate inflammasome function. Understanding these mechanisms is crucial for elucidating the complex regulation of inflammatory responses and the development of therapeutic strategies.The chapter discusses the role of post-translational modifications (PTMs) in regulating NLRP3 inflammasome signaling. PTMs, including phosphorylation, ubiquitination, SUMOylation, acetylation, and glycosylation, influence the activity of inflammasome components through effects on protein-protein interactions, stability, and localization. The review highlights how these modifications can both activate and inhibit inflammasome signaling, contributing to the regulation of inflammatory responses. Key points include: 1. **Non-transcriptional Priming**: Inflammasome activation requires two signals: a priming signal that drives NLRP3 expression and an activating signal that triggers its activation. PTMs like deubiquitination and phosphorylation play crucial roles in this process. 2. **NLRP3 Phosphorylation**: Phosphorylation at specific sites on NLRP3, such as Serine-5 and Serine-198, can inhibit or activate inflammasome activity, respectively. Kinases like AKT, JNK, and BTK are involved in these modifications. 3. **Ubiquitination and SUMOylation**: Ubiquitination at Lys 496 and SUMOylation at Lys 689 can promote or inhibit inflammasome activation, respectively. E3 ligases like TRIM31 and Cbl-b are involved in these modifications. 4. **Palmitoylation**: Palmitoylation can affect NLRP3 stability and protein-protein interactions, with zDHHC12 promoting degradation and zDHHC5 enhancing binding to NEK7. 5. **Other Modifications**: Acetylation, nitrosylation, and ADP-ribosylation also influence inflammasome activity, with specific modifications affecting cytokine release and cell death. 6. **Adaptor Protein ASC**: PTMs on ASC, such as phosphorylation and ubiquitination, regulate speck formation and inflammasome activation. 7. **Caspase-1**: Caspase-1 is modified by phosphorylation, ubiquitination, and nitrosylation, each influencing its activity and downstream signaling. 8. **Gasdermin D**: Gasdermin D is modified by phosphorylation, succination, palmitoylation, and oxidation, each contributing to its pore-forming and pyroptotic functions. 9. **IL-1β**: Pro-IL-1β is modified by glutathionylation and ubiquitination, affecting its stability and activity. The review concludes by highlighting outstanding questions about the integration of PTMs, the dominance of specific modifications, and the cooperative signaling pathways that regulate inflammasome function. Understanding these mechanisms is crucial for elucidating the complex regulation of inflammatory responses and the development of therapeutic strategies.