Gasdermin D Signaling & Inflammasome Research

Pyroptosis is an inflammatory programmed cell death that is initiated in response to pathogen- or host-derived perturbations of the cytosol. In human and mice pyroptosis is induced by inflammatory caspases, such as caspase-1 and -11 that are activated by inflammasomes. Gasdermin D (GSDMD) is a central mediator of pyroptotic cell death. It contains a functional N-terminal domain and an inhibitory C-terminal domain. Upon caspase-1/11 cleavage of the linker located between the two domains of Gasdermin D, the cleaved N-terminal fragment of Gasdermin D oligomerizes and forms pores on the host cell membrane, leading to cell death called pyroptosis.

AdipoGen Life Sciences is the leading manufacturer and supplier of inflammasome signaling research reagents. The validated high quality inflammasome research tools are used and published daily by the experts in the inflammasome research field.

A New Tool to Measure Pyroptosis:

Gasdermin D (mouse) ELISA Kit

The Gasdermin D (mouse) ELISA Kit (Prod. No. AG-45B-0011) is a sandwich ELISA for quantitative determination of mouse Gasdermin D in cell culture supernatants and in cell extracts. This ELISA is specific for the measurement of natural and recombinant mouse Gasdermin D (full-length and C-terminus cleaved fragment). It does not detect human Gasdermin D.

Cat. No.Product NameProduct Description
AG-45B-0011
Gasdermin D (mouse) ELISA Kit
Detects full-length and cleaved C-terminal mouse Gasdermin D in cell culture supernatants and cell extracts. Does not cross-react with human Gasdermin D.

Gasdermin D Signaling Pathways

Inflammasomes are multimeric protein complexes that comprise a sensor (e.g. NLRP3), an adaptor (ASC/Pycard) and a protease (pro-caspase-1) (1). An inflammasome assembles in response to a diverse range of pathogen-associated or danger-associated molecular patterns (PAMPs or DAMPs), or perturbations in cytoplasmic homeostasis (term "homeostasis-altering molecular processes" (HAMPs)) (2). The inflammasome platform leads to activation of caspase-1, which further induces maturation of interleukin-1β and -18 (IL-1β and IL-18) through proteolytic cleavage of pro-IL-1β and pro-IL-18. Activated caspase-1, and also the recently characterized caspase-11 non-canonical inflammasome pathway, cleave the newly discovered intracellular protein Gasdermin D (3, 4). The Gasdermin family members contain N-terminal domains that are capable of forming membrane pores, whereas the C-terminal domains of Gasdermins function as inhibitors of such cytolysis through intramolecular domain association. Caspase-1 or -11 cleavage of Gasdermin D is required for regulation of Pyroptosis: upon caspase-1/11 cleavage of the Gasdermin N- and C-domain linker, the cleaved N-terminal fragment of Gasdermin D oligomerizes and forms pores on the host cell membrane (5), leading to a cell death called pyroptosis and further activation of inflammasomes by triggering K+ efflux (6). Gasdermin D forming pores regulate the non-conventional secretion of cytokines such as IL-1β, in response to cytosolic LPS and other activators of the inflammasome (7). Neutrophil extrusion of neutrophil extracellular traps (NETs) and concomitant cell death (NETosis), a particular neutrophil defense against pathogens, are dependent on Gasdermin D cleavage by caspase-11 (8). Gasdermin D-mediated pyroptosis is regulated at the level of lipid peroxidation (9) and seems to be a key effector in the LPS-induced lethal sepsis (10).

Caspase-8, an upstream activator of caspase-3, controls apoptotic cell death and prevents RIPK3–MLKL-dependent necroptosis. Caspase-8, activated by the Yersinia effector protein YopJ, also triggers Gasdermin D processing and cell death with different Yersinia species (11).

After formation of the pore at the cellular membrane by Gasdermin D N-terminal fragment, the role and fate of the C-terminus fragment of Gasdermin D is still unclear. Using the Gasdermin D (mouse) ELISA Kit (Prod. No. AG-45B-0011), that detects the C-terminal part of Gasdermin D (as well as the full-length protein), a signal is detected in the supernatant of cells dying by pyroptosis, suggesting that the C-terminal fragment is released from cells, either by chance due to the presence of a pore or for a specific task not yet clear.


LITERATURE REFERENCES:
1. The inflammasome: a molecular platform triggering activation of inflammatory caspases and processing of proILbeta: F. Martinon, et al.; Mol. Cell 10, 417 (2002)

2. Homeostasis-altering molecular processes as mechanisms of inflammasome activation: A. Liston & S.L. Masters; Nat. Rev. Immunol. 17, 208 (2017)

3. Caspase-11 cleaves gasdermin D for noncanonical inflammasome signalling: N. Kayagaki, et al.; Nature 526, 666 (2015)

4. Mechanisms of Gasdermin Family Members in Inflammasome Signaling and Cell Death: S. Feng, et al.; J. Mol. Biol. 430, 3068 (2018)

5. Inflammasome-activated gasdermin D causes pyroptosis by forming membrane pores: X. Liu, et al.; Nature 535, 153 (2016)

6. Gasdermin D Restrains Type I Interferon Response to Cytosolic DNA by Disrupting Ionic Homeostasis: I. Banerjee, et al.; Immunity 49, 413 (2018)

7. The Pore-Forming Protein Gasdermin D Regulates Interleukin-1 Secretion from Living Macrophages: C.L. Evavold, et al.; Immunity 48, 35 (2018)

8. Noncanonical inflammasome signaling elicits gasdermin D-dependent neutrophil extracellular traps: K.W. Chen, et al.; Sci. Immunol. 3, 26 (2018)

9. Lipid Peroxidation Drives Gasdermin D-mediated Pyroptosis in Lethal Polymicrobial Sepsis: R. Kang, et al.; Cell Host Microbe 24, 97 (2018)

10. Chemical disruption of the pyroptotic pore-forming protein gasdermin D inhibits inflammatory cell death and sepsis: J.K. Rathkey, et al.; Sci. Immunol. 3, 26 (2018)

11. Pathogen blockade of TAK1 triggers caspase-8-dependent cleavage of gasdermin D and cell death: P. Orning, et al.; Science (Epub ahead of print) 26 (2018)

The Standard Inflammasome Research Antibodies

Cat. No.Product NameProduct Description
AG-20B-0042
anti-Caspase-1 (p20) (mouse), mAb (Casper-1)Recognizes endogenous full-length and activated (p20 fragment) mouse caspase-1.
AG-20B-0044
anti-Caspase-1 (p10) (mouse), mAb (Casper-2)Recognizes endogenous full-length and activated (p10 fragment) mouse caspase-1.
AG-20B-0048
anti-Caspase-1 (p20) (human), mAb (Bally-1)Recognizes endogenous full-length and activated (p20 fragment) human caspase-1.
AG-20B-0014
anti-NLRP3/NALP3, mAb (Cryo-2)Recognizes mouse and human NLRP3/NALP3.
AG-25B-0006
anti-Asc, pAb (AL177)Recognizes human and mouse Asc.
AG-20B-0064
anti-IL-1alpha (p18) (mouse), mAb (Teo-1)Recognizes full-length and cleaved p18 fragment of mouse IL-1α.
AG-20B-0057
anti-Caspase-8 (human), mAb (C15)Recognizes the p18 subunit of human caspase-8.
AG-20T-0137
anti-Caspase-8 (mouse), mAb (1G12)Recognizes full-length and the p18 cleaved fragment of mouse caspase-8.
AG-20T-0138
anti-Caspase-8 (mouse), mAb (3B10)Recognizes full-length and the p18 cleaved fragment of mouse caspase-8.

Key Inflammasome Activators and Inhibitors

Cat. No.Product NameProduct Description
AG-CR1-3950
Monosodium urate (crystals) [1198-77-2]Potent NLRP3 inflammasome activator.
AG-CR1-3951Monosodium urate (ready-to-use) [1198-77-2]Potent NLRP3 inflammasome activator.
AG-CN2-0020
Nigericin . sodium salt [28643-80-3]Potent NLRP3 inflammasome activator.
AG-CN2-0489
N-Acetyl-D-glucosamine [7512-17-6]Acts as activator of NLRP3 inflammasome by dissociating the enzyme hexokinase from the mitochondria.
AG-CR1-3615
MCC950 . sodium salt [256373-96-3]Potent and selective NLRP3 inflammasome inhibitor.
AG-CN2-0459
Isoliquiritigenin [961-29-5]Inhibits NLRP3-activated Asc oligomerization. Blocks priming and activation steps.
AG-CR1-0013
BAY 11-7082 [19542-67-7]Reduces ATPase activity of the NLRP3 inflammasome.
AG-CR1-3616
(R)-3-Hydroxybutyric acid [625-72-9]Prevents K+-efflux and consequently reduce Asc oligomerization and speck formation.
AG-CR1-3617
(S)-3-Hydroxybutyric acid [6168-83-8]
CDX-H0080
DL-3-Hydroxybutyric acid sodium salt [150-83-4]
AG-CR1-0158
K777 [K11777] [233277-99-1]Broad-range cathepsin inhibitor useful for inflammasome inhibition.

Unique Gasdermin D (mouse-specific) Antibody

Cat. No.Product NameProduct Description
AG-25B-0036
anti-Gasdermin D (mouse), pAb (IN110)Recognizes full-length and cleaved C-terminus domain of mouse gasdermin D. Does not cross-react with human gasdermin D.

Gasdermin D-induced Pyroptosis Inhibitors

Cat. No.Product NameProduct Description
AG-CR1-3698
U-73122 [112648-68-7]U-73122 is a potent cell permeable phospholipase C (PLC) and SERCA inhibitor that has recently been shown to be a useful inflammasome research reagent by acting as a Gasdermin D N-terminal fragment (GSDMD-N)-induced pyroptosis inhibitor. It protects against GSDMD-N cytotoxicity in macrophages or against lethal infection in mice.
AG-3705
Necrosulfonamide
Necrosulfonamide is a potent inhibitor of necroptosis, by blocking the mixed lineage kinase domain-like protein (MLKL) has recently been shown to act as an inhibitor of Gasdermin D that works well for mice studies. It binds directly to Gasdermin D and inhibits the oligomerization of the N-terminus and therefore the pore formation and pyroptosis.



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Brands: AdipoGen Life Sciences, Chemodex
Product groups: Antibodies, Assays, Chemicals