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Biochemical and microscopic analysis of inflammasome complex formation
Umeå University, Faculty of Medicine, Molecular Infection Medicine Sweden (MIMS).
Umeå University, Faculty of Medicine, Molecular Infection Medicine Sweden (MIMS). Department of Molecular Biosciences, The Wenner-Gren Institute, Stockholm University, Sweden.ORCID iD: 0000-0002-1269-8288
Umeå University, Faculty of Medicine, Department of Molecular Biology (Faculty of Medicine). (Erttmann)ORCID iD: 0000-0003-2398-8405
2019 (English)In: DNA Sensors and Inflammasomes / [ed] Jungsan Sohn, Elsevier, 2019, Vol. 625, p. 287-298, article id S0076-6879(19)30173-9Chapter in book (Refereed)
Abstract [en]

Inflammasomes are multiprotein signaling platforms responsible for the maturation of pro-IL-1β and pro-IL-18 as well as the induction of an inflammatory cell death termed pyroptosis. Most inflammasomes consist of an upstream sensor, in most cases an adaptor protein (ASC) and inflammatory caspases such as caspase-1. Upon activation, sensor proteins oligomerize with adaptor proteins, forming large complexes called specks. These complexes can be stabilized and detected by Western blotting or fluorescence microscopy providing a direct evidence of inflammasome activation. Here we describe protocols for two complementary methods for detecting inflammasome complexes: (1) biochemical isolation and detection of ASC oligomers by Western blot analysis and (2) microscopic visualization of active caspase-1—ASC complexes. These protocols have successfully been applied in our recent study to unveil new regulatory mechanisms for different inflammasomes including the DNA sensor AIM2 (Erttmann et al., 2016).

Place, publisher, year, edition, pages
Elsevier, 2019. Vol. 625, p. 287-298, article id S0076-6879(19)30173-9
Series
Methods in Enzymology, ISSN 0076-6879 ; 625
Keywords [en]
Inflammasome complex, ASC specks, Caspase 1, AIM2, IL-1β, Innate immunity
National Category
Cell and Molecular Biology
Research subject
Immunology; cell research; Infectious Diseases
Identifiers
URN: urn:nbn:se:umu:diva-162825DOI: 10.1016/bs.mie.2019.05.014PubMedID: 31455532Scopus ID: 2-s2.0-85067031234ISBN: 978-0-12-818359-5 (print)OAI: oai:DiVA.org:umu-162825DiVA, id: diva2:1347072
Available from: 2019-08-30 Created: 2019-08-30 Last updated: 2019-09-03Bibliographically approved

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Swacha, PatrycjaGekara, Nelson O.Erttmann, Saskia F.

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Swacha, PatrycjaGekara, Nelson O.Erttmann, Saskia F.
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Molecular Infection Medicine Sweden (MIMS)Department of Molecular Biology (Faculty of Medicine)
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