Posttranslational and Therapeutic Control of Gasdermin-Mediated Pyroptosis and Inflammation

Pyroptosis is a proinflammatory form of cell death, mediated by membrane pore-forming proteins called gasdermins. Gasdermin pores allow the release of the pro-inflammatory cytokines IL-1β and IL-18 and cause cell swelling and cell lysis leading to release of other intracellular proteins that act as alarmins to perpetuate inflammation. The best characterized, gasdermin D, forms pores via its N-terminal domain, generated after the cleavage of full length gasdermin D by caspase-1 or -11 (caspase-4/5 in humans) typically upon sensing of intracellular pathogens. Thus, gasdermins were originally thought to largely contribute to pathogen-induced inflammation. We now know that gasdermin family members can also be cleaved by other proteases, such as caspase-3, caspase-8 and granzymes, and that they contribute to sterile inflammation as well as inflammation in autoinflammatory diseases or during cancer immunotherapy. Here we briefly review how and when gasdermin pores are formed, and then focus on emerging endogenous mechanisms and therapeutic approaches that could be used to control pore formation, pyroptosis and downstream inflammation.

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Potent antitumor activity of a glutamyltransferase-derived peptide via an activation of oncosis pathway

Scientific Reports ◽

10.1038/s41598-021-93055-5 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Cheng Fang ◽

Wenhui Li ◽

Ruozhe Yin ◽

Donglie Zhu ◽

Xing Liu ◽

...

Keyword(s):

Antitumor Activity ◽

Pore Formation ◽

Cell Swelling ◽

The Novel ◽

Membrane Pore ◽

Membrane Rupture ◽

Analog Peptide ◽

Dependent Cell ◽

Molecular Components ◽

Higher Sensitivity

AbstractHepatocellular carcinoma (HCC) still presents poor prognosis with high mortality rate, despite of the improvement in the management. The challenge for precision treatment was due to the fact that little targeted therapeutics are available for HCC. Recent studies show that metabolic and circulating peptides serve as endogenous switches for correcting aberrant cellular plasticity. Here we explored the antitumor activity of low molecular components in human umbilical serum and identified a high abundance peptide VI-13 by peptidome analysis, which was recognized as the part of glutamyltransferase signal peptide. We modified VI-13 by inserting four arginines and obtained an analog peptide VI-17 to improve its solubility. Our analyses showed that the peptide VI-17 induced rapid context-dependent cell death, and exhibited a higher sensitivity on hepatoma cells, which is attenuated by polyethylene glycol but not necrotic inhibitors such as z-VAD-fmk or necrostatin-1. Morphologically, VI-17 induced cell swelling, blebbing and membrane rupture with release of cellular ATP and LDH into extracellular media, which is hallmark of oncotic process. Mechanistically, VI-17 induced cell membrane pore formation, degradation of α-tubulin via influx of calcium ion. These results indicated that the novel peptide VI-17 induced oncosis in HCC cells, which could serve as a promising lead for development of therapeutic intervention of HCC.

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Stepwise visualization of membrane pore formation by suilysin, a bacterial cholesterol-dependent cytolysin

eLife ◽

10.7554/elife.04247 ◽

2014 ◽

Vol 3 ◽

Cited By ~ 92

Author(s):

Carl Leung ◽

Natalya V Dudkina ◽

Natalya Lukoyanova ◽

Adrian W Hodel ◽

Irene Farabella ◽

...

Keyword(s):

Pore Formation ◽

Pore Diameter ◽

Membrane Attack Complex ◽

Membrane Insertion ◽

Subunit Structure ◽

Membrane Pore ◽

Force Microscopy ◽

Atomic Force ◽

Pore Forming Proteins ◽

Assembly Pathways

Membrane attack complex/perforin/cholesterol-dependent cytolysin (MACPF/CDC) proteins constitute a major superfamily of pore-forming proteins that act as bacterial virulence factors and effectors in immune defence. Upon binding to the membrane, they convert from the soluble monomeric form to oligomeric, membrane-inserted pores. Using real-time atomic force microscopy (AFM), electron microscopy (EM), and atomic structure fitting, we have mapped the structure and assembly pathways of a bacterial CDC in unprecedented detail and accuracy, focussing on suilysin from Streptococcus suis. We show that suilysin assembly is a noncooperative process that is terminated before the protein inserts into the membrane. The resulting ring-shaped pores and kinetically trapped arc-shaped assemblies are all seen to perforate the membrane, as also visible by the ejection of its lipids. Membrane insertion requires a concerted conformational change of the monomeric subunits, with a marked expansion in pore diameter due to large changes in subunit structure and packing.

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Potentiation of Apoptin-Induced Apoptosis by Cecropin B-Like Antibacterial Peptide ABPs1 in Human HeLa Cervical Cancer Cell Lines is Associated with Membrane Pore Formation and Caspase-3 Activation

Journal of Microbiology and Biotechnology ◽

10.4014/jmb.1209.09076 ◽

2014 ◽

Vol 24 (6) ◽

pp. 756-764 ◽

Cited By ~ 5

Author(s):

Basse Mame Birame ◽

Wang Jigui ◽

Yu Fuxian ◽

Sun Jiazeng ◽

Li Zhili ◽

...

Keyword(s):

Cervical Cancer ◽

Cell Lines ◽

Cancer Cell ◽

Pore Formation ◽

Caspase 3 ◽

Antibacterial Peptide ◽

Cervical Cancer Cell ◽

Membrane Pore ◽

Induced Apoptosis ◽

Hela Cervical Cancer Cell

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Membrane pore formation at protein–lipid interfaces

Trends in Biochemical Sciences ◽

10.1016/j.tibs.2014.09.002 ◽

2014 ◽

Vol 39 (11) ◽

pp. 510-516 ◽

Cited By ~ 95

Author(s):

Robert J.C. Gilbert ◽

Mauro Dalla Serra ◽

Christopher J. Froelich ◽

Mark I. Wallace ◽

Gregor Anderluh

Keyword(s):

Pore Formation ◽

Membrane Pore

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Molecular Basis for Membrane Pore Formation by Bax Protein Carboxyl Terminus

Biochemistry ◽

10.1021/bi301195f ◽

2012 ◽

Vol 51 (46) ◽

pp. 9406-9419 ◽

Cited By ~ 15

Author(s):

Suren A. Tatulian ◽

Pranav Garg ◽

Kathleen N. Nemec ◽

Bo Chen ◽

Annette R. Khaled

Keyword(s):

Molecular Basis ◽

Pore Formation ◽

Carboxyl Terminus ◽

Membrane Pore ◽

Bax Protein

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Effects of Onchung-eum, an Herbal Prescription, on 5-Fluorouracil–Induced Oral Mucositis

Integrative Cancer Therapies ◽

10.1177/1534735418805560 ◽

2018 ◽

Vol 17 (4) ◽

pp. 1285-1296

Author(s):

Jae-Woo Park ◽

Jayoung Oh ◽

Seok-Jae Ko ◽

Mun Seog Chang ◽

Jinsung Kim

Keyword(s):

Inflammatory Cytokines ◽

Oral Mucositis ◽

Caspase 3 ◽

Skin Diseases ◽

Nuclear Factor Κb ◽

Control Group ◽

Nuclear Factor ◽

Cheek Pouches ◽

Herbal Prescription ◽

Pro Inflammatory Cytokines

In most cancer patients, chemotherapy-induced oral mucositis (OM) is a frequent side effect, leading to low quality of life and delay in therapy. The aim of this study was to evaluate the effects of Onchung-eum, a well-known herbal prescription in traditional medicine comprising 8 herbs that has long been used for skin diseases, on 5-fluorouracil (5-FU)–induced OM in human pharyngeal cells and golden Syrian hamsters. DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging activity, MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay, and reactive oxygen species production were measured in vitro. The effects of Onchung-eum on OM of hamster cheek pouches induced by 5-FU were evaluated histologically and using TUNEL assay. In addition, the expression of nuclear factor-κB, caspase-3, and pro-inflammatory cytokines were measured by immunoblotting and immunohistochemistry. Significantly increased cell viability was observed in the Onchung-eum–treated groups compared with the 5-FU–treated control group. In 500 and 1000 mg/kg Onchung-eum–treated groups, the damaged epithelial layers in the cheek pouches of hamsters were significantly recovered. Moreover, at all concentrations, cell death in the cheek pouches of hamsters in the Onchung-eum–treated groups significantly decreased. The expression of pro-inflammatory cytokines, nuclear factor-κB, and caspase-3 also significantly decreased in Onchung-eum–treated groups at 500 and 1000 mg/kg. In conclusion, this study revealed that Onchung-eum can be used to treat chemotherapy-induced OM. However, further studies are required to understand the underlying mechanisms.

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Membrane pore formation by the human immunodeficiency virus type-1 neutralizing anti-gp41 antibody 2F5

Chemistry and Physics of Lipids ◽

10.1016/j.chemphyslip.2010.05.138 ◽

2010 ◽

Vol 163 ◽

pp. S46

Author(s):

Rubén Maeso ◽

Renate Kunert ◽

José L. Nieva

Keyword(s):

Human Immunodeficiency Virus ◽

Human Immunodeficiency Virus Type ◽

Virus Type ◽

Pore Formation ◽

Membrane Pore ◽

Immunodeficiency Virus

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Genetically-Engineered Protease-Activated Triggers in a Pore-Forming Protein

MRS Proceedings ◽

10.1557/proc-330-209 ◽

1993 ◽

Vol 330 ◽

Cited By ~ 1

Author(s):

Barbara Walker ◽

Nathan Walsh ◽

Hagan Bayley

Keyword(s):

Genetic Engineering ◽

Cancer Cells ◽

Pore Formation ◽

Polypeptide Chain ◽

Genetically Engineered ◽

Recognition Sequence ◽

Selective Activation ◽

Selective Destruction ◽

Pore Forming Proteins ◽

Central Loop

ABSTRACTProtease-activated triggers have been introduced Into a pore-forming protein, staphylococcal a-hemolysin (αHL). The hemolysin was remodeled by genetic engineering to form two-chain constructs with redundant polypeptide sequences at the central loop, the Integrity of which Is crucial for efficient pore formation. The new hemolysins are activated when the polypeptide extensions are removed by proteases. By alterating the protease recognition sequence in the loop, selective activation by specified proteases can be obtained. Protease-triggered pore-forming proteins might be used for the selective destruction of cancer cells that bear tumor-associated proteases. When certain two-chain constructs are treated with proteases, a full-length polypeptide chain forms as the result of a protease-mediated transpeptidation reaction. This reaction might be used to produce chimeric hemolysins that are Inaccessible by conventional routes.

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