An Assay for the Seeding of Homotypic Pyrin Domain Filament Transitions

Inga V Hochheiser; Matthias Geyer

doi:10.1007/978-1-0716-2449-4_13

An Assay for the Seeding of Homotypic Pyrin Domain Filament Transitions

Methods Mol Biol. 2022:2523:197-207. doi: 10.1007/978-1-0716-2449-4_13.

Authors

Inga V Hochheiser¹, Matthias Geyer²

Affiliations

¹ Institute of Structural Biology, University Hospital Bonn, University of Bonn, Bonn, Germany.
² Institute of Structural Biology, University Hospital Bonn, University of Bonn, Bonn, Germany. matthias.geyer@uni-bonn.de.

PMID: 35759199
DOI: 10.1007/978-1-0716-2449-4_13

Abstract

Pattern recognition receptors of innate immune cells allow the recognition of invariant microbial structures. The nucleotide-binding oligomerization domain-like receptors (NLRs) comprise 22 members, divided into 3 subfamilies. Homotypic pyrin domain (PYD) interactions were shown to mediate the interaction of inflammasome forming NLRPs with the adaptor protein ASC, bridging the interaction to caspase-1 and resulting in caspase-1-induced cytokine maturation and pyroptotic cell death. Here we describe a NLRP3^PYD-mediated ASC polymerization assay that reconstitutes the transition from the NLRP3^PYD nucleation seed to ASC adaptor filament elongation with recombinant proteins.

Keywords: ASC; Electron microscopy; Filament elongation; Filament nucleation; Inflammasome; Innate immunity; NLRP3; PYD; Pyrin domain.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Caspase 1 / metabolism
Cytoskeletal Proteins / metabolism
NLR Family, Pyrin Domain-Containing 3 Protein* / metabolism
Protein Binding
Pyrin Domain*

Substances

Cytoskeletal Proteins
NLR Family, Pyrin Domain-Containing 3 Protein
Caspase 1