A single domain antibody fragment that recognizes the adaptor ASC defines the role of ASC domains in inflammasome assembly

Myeloid cells assemble inflammasomes in response to infection or cell damage; cytosolic sensors activate pro–caspase-1, indirectly for the most part, via the adaptors ASC and NLRC4. This leads to secretion of proinflammatory cytokines and pyroptosis. To explore complex formation under physiological...

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Autores principales: Schmidt, Florian I., Lu, Alvin, Chen, Jeff W., Ruan, Jianbin, Tang, Catherine, Wu, Hao, Ploegh, Hidde L.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Rockefeller University Press 2016
Materias:
Research Articles
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4854733/
https://www.ncbi.nlm.nih.gov/pubmed/27069117
http://dx.doi.org/10.1084/jem.20151790
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author Schmidt, Florian I.
Lu, Alvin
Chen, Jeff W.
Ruan, Jianbin
Tang, Catherine
Wu, Hao
Ploegh, Hidde L.
author_facet Schmidt, Florian I.
Lu, Alvin
Chen, Jeff W.
Ruan, Jianbin
Tang, Catherine
Wu, Hao
Ploegh, Hidde L.
author_sort Schmidt, Florian I.
collection PubMed
description Myeloid cells assemble inflammasomes in response to infection or cell damage; cytosolic sensors activate pro–caspase-1, indirectly for the most part, via the adaptors ASC and NLRC4. This leads to secretion of proinflammatory cytokines and pyroptosis. To explore complex formation under physiological conditions, we generated an alpaca single domain antibody, VHH(ASC), which specifically recognizes the CARD of human ASC via its type II interface. VHH(ASC) not only impairs ASC(CARD) interactions in vitro, but also inhibits inflammasome activation in response to NLRP3, AIM2, and NAIP triggers when expressed in living cells, highlighting a role of ASC in all three types of inflammasomes. VHH(ASC) leaves the Pyrin domain of ASC functional and stabilizes a filamentous intermediate of inflammasome activation. Incorporation of VHH(ASC)-EGFP into these structures allowed the visualization of endogenous ASC(PYD) filaments for the first time. These data revealed that cross-linking of ASC(PYD) filaments via ASC(CARD) mediates the assembly of ASC foci.
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spelling pubmed-48547332016-11-02 A single domain antibody fragment that recognizes the adaptor ASC defines the role of ASC domains in inflammasome assembly Schmidt, Florian I. Lu, Alvin Chen, Jeff W. Ruan, Jianbin Tang, Catherine Wu, Hao Ploegh, Hidde L. J Exp Med Research Articles Myeloid cells assemble inflammasomes in response to infection or cell damage; cytosolic sensors activate pro–caspase-1, indirectly for the most part, via the adaptors ASC and NLRC4. This leads to secretion of proinflammatory cytokines and pyroptosis. To explore complex formation under physiological conditions, we generated an alpaca single domain antibody, VHH(ASC), which specifically recognizes the CARD of human ASC via its type II interface. VHH(ASC) not only impairs ASC(CARD) interactions in vitro, but also inhibits inflammasome activation in response to NLRP3, AIM2, and NAIP triggers when expressed in living cells, highlighting a role of ASC in all three types of inflammasomes. VHH(ASC) leaves the Pyrin domain of ASC functional and stabilizes a filamentous intermediate of inflammasome activation. Incorporation of VHH(ASC)-EGFP into these structures allowed the visualization of endogenous ASC(PYD) filaments for the first time. These data revealed that cross-linking of ASC(PYD) filaments via ASC(CARD) mediates the assembly of ASC foci. The Rockefeller University Press 2016-05-02 /pmc/articles/PMC4854733/ /pubmed/27069117 http://dx.doi.org/10.1084/jem.20151790 Text en © 2016 Schmidt et al. This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).
spellingShingle Research Articles
Schmidt, Florian I.
Lu, Alvin
Chen, Jeff W.
Ruan, Jianbin
Tang, Catherine
Wu, Hao
Ploegh, Hidde L.
A single domain antibody fragment that recognizes the adaptor ASC defines the role of ASC domains in inflammasome assembly
title A single domain antibody fragment that recognizes the adaptor ASC defines the role of ASC domains in inflammasome assembly
title_full A single domain antibody fragment that recognizes the adaptor ASC defines the role of ASC domains in inflammasome assembly
title_fullStr A single domain antibody fragment that recognizes the adaptor ASC defines the role of ASC domains in inflammasome assembly
title_full_unstemmed A single domain antibody fragment that recognizes the adaptor ASC defines the role of ASC domains in inflammasome assembly
title_short A single domain antibody fragment that recognizes the adaptor ASC defines the role of ASC domains in inflammasome assembly
title_sort single domain antibody fragment that recognizes the adaptor asc defines the role of asc domains in inflammasome assembly
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4854733/
https://www.ncbi.nlm.nih.gov/pubmed/27069117
http://dx.doi.org/10.1084/jem.20151790
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