DNA Sensors and Inflammasomes

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. Intercellular communication in the innate immune system through the cGAS-STING pathway Ramya Nandakumar, Samuel Joseph Windross and Søren R. Paludan 2. Analysis of human cGAS activity and structure Wen Zhou, Aaron T. Whiteley and Philip J. Kranzusch 3. Chemical synthesis, purification, and characterization of 3′-5′-linked canonical cyclic dinucleotides (CDNs) Changhao Wang, Min Hao, Qianqian Qi, Yashao Chen and Jörg S. Hartig 4. An SPR-based analysis of cGAS substrate K_D and steady-state K_M values Erik C. Ralph and Justin Hall 5. A pyrophosphatase-coupled assay to monitor the NTase activity of cGAS Richard Hooy and Jungsan Sohn 6. Tracking the polymerization of DNA sensors and inflammasomes using FRET Zachary Mazanek and Jungsan Sohn 7. Monitoring gasdermin pore formation in vitro Shiyu Xia, Jianbin Ruan and Hao Wu 8. Measuring TREX1 and TREX2 exonuclease activities Wayne O. Hemphill and Fred W. Perrino 9. Kinetics of DNA: Protein association and dissociation by stopped-flow spectroscopy Johannes Rudolph and Karolin Luger 10. Characterization of DNA bound cyclic GMP-AMP synthase using atomic force microscopy imaging Alexander Lushnikov, Richard Hooy, Jungsan Sohn and Alexey Krasnoslobodtsev 11. Preparation of filamentous proteins for electron microscopy visualization and reconstruction Mariusz Matyszewski and Jungsan Sohn 12. Cryo-EM studies of NAIP–NLRC4 inflammasomes Nicole Haloupek, Patricia Grob, Jeannette Tenthorey, Russell E. Vance and Eva Nogales 13. An integrative protocol for the structure determination of the mouse ASC-PYD filament Adam Mazur, Petr Broz and Sebastian Hiller 14. Protein interactions of the inflammasome adapter ASC by solution NMR Pedro Diaz-Parga and Eva de Alba 15. Tracking DNA sensor and inflammasome complexes in cells using confocal microscopy Brendan Antiochos 16. Assessment of inflammasome and type I IFN responses to DNA viruses and DNA PAMPS Guido Massaccesi and Michael A. Chattergoon 17. Biochemical and microscopic analysis of inflammasome complex formation Patrycja Swacha, Nelson Ong’ondo Gekara and Saskia F. Erttmann 18. Comet and micronucleus assays for analyzing DNA damage and genome integrity Hui Jiang and Nelson Ong'ondo Gekara 19. Workflows and considerations for investigating protein interactions of viral DNA sensors Timothy R. Howard, Bokai Song and Ileana M. Cristea 20. TUBE and UbiCRest assays for elucidating polyubiquitin modifications in protein complexes Swarupa Panda and Nelson Ong'ondo Gekara