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.
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 KD and steady-state KM
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
Jungsan Sohn is at Johns Hopkins School of Medicine, MD, USA
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