DNA Sensors and Inflammasomes, Volume 625, the latest release in the Methods in Enzymology series, continues the legacy of this premier serial with quality chapters authored by leaders in the field. New sections in this release include Phosphorylation and dimerization of STING and IRF3, cGAS enzymology, Synthesis and identification of immuno-stimulatory CDNs, Tracking cGAS activity/ cGAMP formation using SPR/NMR, Using an enzyme coupled assay to track cGAS activity under steady states, Tracking the polymerization of DNA sensors, inflammasome receptors, and downstream signaling partners using FRET, NLRC4 structure, Tracking TREX1 activity, DNA association and dissociation kinetics of PARP1, and more.
1. Cover image - p. iv 2. Title page - p. iv 3. Table of Contents - p. iv 4. Copyright - p. iv 5. Contributors - p. xiii 6. Preface - p. xvii 7. Chapter One: Intercellular communication in the innate immune system through the cGAS-STING pathway - p. xvii 8. Chapter Two: Analysis of human cGAS activity and structure - p. 11 9. Chapter Three: Chemical synthesis, purification, and characterization of 3′-5′-linked canonical cyclic dinucleotides (CDNs) - p. 40 10. Chapter Four: An SPR-based analysis of cGAS substrate KD and steady-state KM values - p. 59 11. Chapter Five: A pyrophosphatase-coupled assay to monitor the NTase activity of cGAS - p. 76 12. Chapter Six: Tracking the polymerization of DNA sensors and inflammasomes using FRET - p. 86 13. Chapter Seven: Monitoring gasdermin pore formation in vitro - p. 94 14. Chapter Eight: Measuring TREX1 and TREX2 exonuclease activities - p. 107 15. Chapter Nine: Kinetics of DNA–protein association and dissociation by stopped-flow spectroscopy - p. 133 16. Chapter Ten: Characterization of DNA bound cyclic GMP-AMP synthase using atomic force microscopy imaging - p. 156 17. Chapter Eleven: Preparation of filamentous proteins for electron microscopy visualization and reconstruction - p. 166 18. Chapter Twelve: Cryo-EM studies of NAIP–NLRC4 inflammasomes - p. 176 19. Chapter Thirteen: An integrative protocol for the structure determination of the mouse ASC-PYD filament - p. 204 20. Chapter Fourteen: Protein interactions of the inflammasome adapter ASC by solution NMR - p. 222 21. Chapter Fifteen: Tracking DNA sensor and inflammasome complexes in cells using confocal microscopy - p. 252 22. Chapter Sixteen: Assessment of inflammasome and type I IFN responses to DNA viruses and DNA PAMPS - p. 267 23. Chapter Seventeen: Biochemical and microscopic analysis of inflammasome complex formation - p. 285 24. Chapter Eighteen: Comet and micronucleus assays for analyzing DNA damage and genome integrity - p. 298 25. Chapter Nineteen: Workflows and considerations for investigating protein interactions of viral DNA sensors - p. 307 26. Chapter Twenty: TUBE and UbiCRest assays for elucidating polyubiquitin modifications in protein complexes - p. 338
DNA Sensors and Inflammasomes, Volume 625, the latest release in the Methods in Enzymology series, continues the legacy of this premier serial with quality chapters authored by leaders in the field. New sections in this release include Phosphorylation and dimerization of STING and IRF3, cGAS enzymology, Synthesis and identification of immuno-stimulatory CDNs, Tracking cGAS activity/ cGAMP formation using SPR/NMR, Using an enzyme coupled assay to track cGAS activity under steady states, Tracking the polymerization of DNA sensors, inflammasome receptors, and downstream signaling partners using FRET, NLRC4 structure, Tracking TREX1 activity, DNA association and dissociation kinetics of PARP1, and more.
1. Cover image - p. iv 2. Title page - p. iv 3. Table of Contents - p. iv 4. Copyright - p. iv 5. Contributors - p. xiii 6. Preface - p. xvii 7. Chapter One: Intercellular communication in the innate immune system through the cGAS-STING pathway - p. xvii 8. Chapter Two: Analysis of human cGAS activity and structure - p. 11 9. Chapter Three: Chemical synthesis, purification, and characterization of 3′-5′-linked canonical cyclic dinucleotides (CDNs) - p. 40 10. Chapter Four: An SPR-based analysis of cGAS substrate KD and steady-state KM values - p. 59 11. Chapter Five: A pyrophosphatase-coupled assay to monitor the NTase activity of cGAS - p. 76 12. Chapter Six: Tracking the polymerization of DNA sensors and inflammasomes using FRET - p. 86 13. Chapter Seven: Monitoring gasdermin pore formation in vitro - p. 94 14. Chapter Eight: Measuring TREX1 and TREX2 exonuclease activities - p. 107 15. Chapter Nine: Kinetics of DNA–protein association and dissociation by stopped-flow spectroscopy - p. 133 16. Chapter Ten: Characterization of DNA bound cyclic GMP-AMP synthase using atomic force microscopy imaging - p. 156 17. Chapter Eleven: Preparation of filamentous proteins for electron microscopy visualization and reconstruction - p. 166 18. Chapter Twelve: Cryo-EM studies of NAIP–NLRC4 inflammasomes - p. 176 19. Chapter Thirteen: An integrative protocol for the structure determination of the mouse ASC-PYD filament - p. 204 20. Chapter Fourteen: Protein interactions of the inflammasome adapter ASC by solution NMR - p. 222 21. Chapter Fifteen: Tracking DNA sensor and inflammasome complexes in cells using confocal microscopy - p. 252 22. Chapter Sixteen: Assessment of inflammasome and type I IFN responses to DNA viruses and DNA PAMPS - p. 267 23. Chapter Seventeen: Biochemical and microscopic analysis of inflammasome complex formation - p. 285 24. Chapter Eighteen: Comet and micronucleus assays for analyzing DNA damage and genome integrity - p. 298 25. Chapter Nineteen: Workflows and considerations for investigating protein interactions of viral DNA sensors - p. 307 26. Chapter Twenty: TUBE and UbiCRest assays for elucidating polyubiquitin modifications in protein complexes - p. 338
