Plus de 40 ans d’expérience à votre service

Tel. : 01 45 33 67 17 / E-mail : coger@coger.fr

Stock épuisé.
En rupture de stock
Quantité minimum d'achat
La quantité minimum d'achat n'est pas atteinte

 

 

Inflammasomes and Gasdermin D Signaling Pathways

Gasdermin D Signaling

 

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 proIL-beta: 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 non-canonical 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)
 
 

New Reagents for Gasdermin Research


 

NEW 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.
 

Specificity:

Gasdermin D is tested from supernatants of Bone Marrow-Derived Macrophages cells (BMDMs) transfected with LPS from different knockout mice strains (see Figure 1). Only the supernatants from WT and NLRP3-/- strains contain the protein Gasdermin D. Gasdermin D is also tested from cell extracts (lysed with a Triton X-100 buffer) of Bone Marrow-Derived Macrophages cells from WT and Gasdermin D knockout mice strains (see Figure 2).
   Gasdermin D (mouse) ELISA Kit

 

Download the ELISA Kit Manual

Download the Scientific Poster presented at the EMBO Inflammasome Symposia 2018 in Munich

 


 

NEW anti-Gasdermin D (mouse), pAb (IN110) 

AdipoGen Life Sciences' anti-Gasdermin D (mouse), pAb (IN110) (Prod. No. AG-25B-0036) is a polyclonal antibody immunised with the recombinant C-terminus domain of mouse Gasdermin D. The antibody recognizes full-length and the cleaved C-terminus of mouse Gasdermin D, does not cross-react with human Gasdermin D and works specifically in Western Blot application to detect the cleaved C-terminal Gasdermin D.
 

Specificity:

Figure: Mouse Gasdermin D (full-length and cleaved p22 fragments) are detected by immunoblotting using anti-Gasdermin D (mouse), pAb (IN110) (Prod. No. AG-25B-0036).

Method: Gasdermin D is analyzed by Western blot in cell extracts of bone marrow-derived macrophage cells (BMDMs) (WT, Gasdermin -/- or Asc -/-) treated with LPS (50ng/ml; Prod. No. AG-CU1-0001) for 3h and +/- Nigericin (5μM for 2.5h, Prod. No. AG-CN2-0020). Cell extracts are separated by SDS-PAGE under reducing conditions, transferred to nitrocellulose and incubated with anti-Gasdermin D (mouse), pAb (IN110) (0.5µg/ml). After addition of an anti-guinea pig secondary antibody coupled to HRP (1/5000), proteins are visualized by a chemiluminescence detection system.

Picture courtesy of Prof. Olaf Gross, University Medical Center Freiburg, Germany
   anti-Gasdermin D (mouse), pAb (IN110)
interligne
interligne
Gasdermin D (mouse) ELISA Kit Gasdermin D (mouse) ELISA Kit 96 wells Ref. AG-45B-0011-KI01 Marque ADIPOGEN LIFE SCIENCES 590.00 531.00 H.T.
 
anti-Gasdermin D (mouse), pAb (IN110) anti-Gasdermin D (mouse), pAb (IN110) 100 µg Ref. AG-25B-0036-C100 Marque ADIPOGEN LIFE SCIENCES 360.00 324.00 H.T.
 
interligne
anti-Caspase-1 (p20) (mouse), mAb (Casper-1) anti-Caspase-1 (p20) (mouse), mAb (Casper-1) 100 µg Ref. AG-20B-0042-C100 Marque ADIPOGEN LIFE SCIENCES 360.00 324.00 H.T.
 
interligne
anti-Caspase-1 (p10) (mouse), mAb (Casper-2) anti-Caspase-1 (p10) (mouse), mAb (Casper-2) 100 µg Ref. AG-20B-0044-C100 Marque ADIPOGEN LIFE SCIENCES 360.00 324.00 H.T.
 
interligne
anti-Caspase-1 (p20) (human), mAb (Bally-1) anti-Caspase-1 (p20) (human), mAb (Bally-1) 100 µg Ref. AG-20B-0048-C100 Marque ADIPOGEN LIFE SCIENCES 360.00 324.00 H.T.
 
interligne
anti-NLRP3/NALP3, mAb (Cryo-2) anti-NLRP3/NALP3, mAb (Cryo-2) 100 µg Ref. AG-20B-0014-C100 Marque ADIPOGEN LIFE SCIENCES 280.00 252.00 H.T.
 
anti-Asc, pAb (AL177) anti-Asc, pAb (AL177) 100 µg Ref. AG-25B-0006-C100 Marque ADIPOGEN LIFE SCIENCES 400.00 360.00 H.T.
 
interligne
anti-Asc, pAb (AL177) (preservative free) anti-Asc, pAb (AL177) (preservative free) 100 µg Ref. AG-25B-0006PF-C100 Marque ADIPOGEN LIFE SCIENCES 400.00 360.00 H.T.
 
anti-Asc, pAb (AL177) (ATTO 647N) anti-Asc, pAb (AL177) (ATTO 647N) 100 µg Ref. AG-25B-0006TS-C100 Marque ADIPOGEN LIFE SCIENCES 430.00 387.00 H.T.
 
interligne
anti-IL-1alpha (p18) (mouse), mAb (Teo-1) anti-IL-1alpha (p18) (mouse), mAb (Teo-1) 100 µg Ref. AG-20B-0064-C100 Marque ADIPOGEN LIFE SCIENCES 140.00 126.00 H.T.
 
anti-Caspase-8 (human), mAb (C15) anti-Caspase-8 (human), mAb (C15) 50 µg Ref. AG-20B-0057-C050 Marque ADIPOGEN LIFE SCIENCES 230.00 207.00 H.T.
 
interligne
anti-Caspase-8 (human), mAb (C15) anti-Caspase-8 (human), mAb (C15) 100 µg Ref. AG-20B-0057-C100 Marque ADIPOGEN LIFE SCIENCES 370.00 333.00 H.T.
 
anti-Caspase-8 (mouse), mAb (1G12) anti-Caspase-8 (mouse), mAb (1G12) 100 µg Ref. AG-20T-0137-C100 Marque ADIPOGEN LIFE SCIENCES 280.00 252.00 H.T.
 
interligne
anti-Caspase-8 (mouse), mAb (3B10) anti-Caspase-8 (mouse), mAb (3B10) 100 µg Ref. AG-20T-0138-C100 Marque ADIPOGEN LIFE SCIENCES 280.00 252.00 H.T.
 
Monosodium urate (crystals) Monosodium urate (crystals) 2 mg Ref. AG-CR1-3950-M002 Marque ADIPOGEN LIFE SCIENCES 78.00 70.20 H.T.
 
interligne
Monosodium urate (crystals) Monosodium urate (crystals) 2 x 2 mg Ref. AG-CR1-3950-2002 Marque ADIPOGEN LIFE SCIENCES 120.00 108.00 H.T.
 
Monosodium urate (ready-to-use) Monosodium urate (ready-to-use) 10 mg Ref. AG-CR1-3951-M010 Marque ADIPOGEN LIFE SCIENCES 380.00 342.00 H.T.
 
interligne
Nigericin . sodium salt Nigericin . sodium salt 5 mg Ref. AG-CN2-0020-M005 Marque ADIPOGEN LIFE SCIENCES 55.00 49.50 H.T.
 
Nigericin . sodium salt Nigericin . sodium salt 25 mg Ref. AG-CN2-0020-M025 Marque ADIPOGEN LIFE SCIENCES 165.00 148.50 H.T.
 
interligne
N-Acetyl-D-glucosamine N-Acetyl-D-glucosamine 250 mg Ref. AG-CN2-0489-M250 Marque ADIPOGEN LIFE SCIENCES 10.00 9.00 H.T.
 
N-Acetyl-D-glucosamine N-Acetyl-D-glucosamine 1 g Ref. AG-CN2-0489-G001 Marque ADIPOGEN LIFE SCIENCES 15.00 13.50 H.T.
 
interligne
N-Acetyl-D-glucosamine N-Acetyl-D-glucosamine 5 g Ref. AG-CN2-0489-G005 Marque ADIPOGEN LIFE SCIENCES 45.00 40.50 H.T.
 
MCC950 . sodium salt MCC950 . sodium salt 1 mg Ref. AG-CR1-3615-M001 Marque ADIPOGEN LIFE SCIENCES 45.00 40.50 H.T.
 
interligne
MCC950 . sodium salt MCC950 . sodium salt 5 mg Ref. AG-CR1-3615-M005 Marque ADIPOGEN LIFE SCIENCES 80.00 72.00 H.T.
 
MCC950 . sodium salt MCC950 . sodium salt 10 mg Ref. AG-CR1-3615-M010 Marque ADIPOGEN LIFE SCIENCES 120.00 108.00 H.T.
 
interligne
Isoliquiritigenin Isoliquiritigenin 10 mg Ref. AG-CN2-0459-M010 Marque ADIPOGEN LIFE SCIENCES 120.00 108.00 H.T.
 
Isoliquiritigenin Isoliquiritigenin 50 mg Ref. AG-CN2-0459-M050 Marque ADIPOGEN LIFE SCIENCES 360.00 324.00 H.T.
 
interligne
BAY 11-7082 BAY 11-7082 10 mg Ref. AG-CR1-0013-M010 Marque ADIPOGEN LIFE SCIENCES 70.00 63.00 H.T.
 
BAY 11-7082 BAY 11-7082 50 mg Ref. AG-CR1-0013-M050 Marque ADIPOGEN LIFE SCIENCES 280.00 252.00 H.T.
 
interligne
(R)-3-Hydroxybutyric acid (R)-3-Hydroxybutyric acid 25 mg Ref. AG-CR1-3616-M025 Marque ADIPOGEN LIFE SCIENCES 40.00 36.00 H.T.
 
(R)-3-Hydroxybutyric acid (R)-3-Hydroxybutyric acid 100 mg Ref. AG-CR1-3616-M100 Marque ADIPOGEN LIFE SCIENCES 85.00 76.50 H.T.
 
interligne
(S)-3-Hydroxybutyric acid (S)-3-Hydroxybutyric acid 25 mg Ref. AG-CR1-3617-M025 Marque ADIPOGEN LIFE SCIENCES 35.00 31.50 H.T.
 
(S)-3-Hydroxybutyric acid (S)-3-Hydroxybutyric acid 100 mg Ref. AG-CR1-3617-M100 Marque ADIPOGEN LIFE SCIENCES 80.00 72.00 H.T.
 
interligne interligne
K777 [K11777] K777 [K11777] 1 mg Ref. AG-CR1-0158-M001 Marque ADIPOGEN LIFE SCIENCES 40.00 36.00 H.T.
 
K777 [K11777] K777 [K11777] 5 mg Ref. AG-CR1-0158-M005 Marque ADIPOGEN LIFE SCIENCES 160.00 144.00 H.T.
 
interligne
U-73122 U-73122 5 mg Ref. AG-CR1-3698-M005 Marque ADIPOGEN LIFE SCIENCES 68.00 61.20 H.T.
 
U-73122 U-73122 1 mg Ref. AG-CR1-3698-M001 Marque ADIPOGEN LIFE SCIENCES 17.00 15.30 H.T.
 
interligne