Protocol to purify recombinant inflammatory caspases and assess their catalytic activity in vitro.
Cell Biology
Immunology
Molecular Biology
Protein expression and purification
Journal
STAR protocols
ISSN: 2666-1667
Titre abrégé: STAR Protoc
Pays: United States
ID NLM: 101769501
Informations de publication
Date de publication:
16 12 2022
16 12 2022
Historique:
received:
02
09
2022
revised:
26
09
2022
accepted:
20
10
2022
entrez:
3
1
2023
pubmed:
4
1
2023
medline:
6
1
2023
Statut:
ppublish
Résumé
The inflammatory caspases, such as caspase-1, caspase-4, or caspase-11, are key enzymes in mammalian innate immunity as they control inflammasome-dependent inflammation. Assessing the specific proteolytic activities of these caspases in the context of a cell remains challenging, which is why in vitro studies of their catalytic activity have proven useful. Herein, we describe a detailed protocol for the purification of recombinant inflammatory caspases after heterologous expression in bacteria and how to assess and quantify cleavage of full-length protein substrates. For complete details on the use and execution of this protocol, please refer to Devant et al. (2021).
Identifiants
pubmed: 36595884
pii: S2666-1667(22)00728-6
doi: 10.1016/j.xpro.2022.101848
pmc: PMC9668568
pii:
doi:
Substances chimiques
Caspases
EC 3.4.22.-
Inflammasomes
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Research Support, N.I.H., Extramural
Langues
eng
Sous-ensembles de citation
IM
Pagination
101848Subventions
Organisme : NIDDK NIH HHS
ID : P30 DK034854
Pays : United States
Informations de copyright
Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.
Déclaration de conflit d'intérêts
Declaration of interests J.C.K. consults and holds equity in Corner Therapeutics, Larkspur Biosciences, and Neumora Therapeutics. None of these relationships influenced the manuscript.
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