2. Academic Validation
  3. Clustering of NLRP3 induced by membrane or protein scaffolds promotes inflammasome assembly

Clustering of NLRP3 induced by membrane or protein scaffolds promotes inflammasome assembly

  • Nat Commun. 2025 May 27;16(1):4887. doi: 10.1038/s41467-025-60277-4.
Elvira Boršić 1 2,  Taja Železnik Ramuta 1,  Sara Orehek 1,  Mateja Erdani Kreft 3,  Matthias Geyer 4,  Roman Jerala 1 5,  Iva Hafner-Bratkovič 6 7 8
Affiliations

Affiliations

  • 1 Department of Synthetic Biology and Immunology, National Institute of Chemistry, Ljubljana, Slovenia.
  • 2 Interdisciplinary Doctoral Study of Biomedicine, Faculty of Medicine, University of Ljubljana, Ljubljana, Slovenia.
  • 3 Institute of Cell Biology, Faculty of Medicine, University of Ljubljana, Ljubljana, Slovenia.
  • 4 Institute of Structural Biology, University Clinics Bonn, University of Bonn, Bonn, Germany.
  • 5 Centre for the Technologies of Gene and Cell Therapy, National Institute of Chemistry, Ljubljana, Slovenia.
  • 6 Department of Synthetic Biology and Immunology, National Institute of Chemistry, Ljubljana, Slovenia. [email protected].
  • 7 Institute of Cell Biology, Faculty of Medicine, University of Ljubljana, Ljubljana, Slovenia. [email protected].
  • 8 EN-FIST Centre of Excellence, Ljubljana, SI-1000, Slovenia. [email protected].
PMID: 40425567 DOI: 10.1038/s41467-025-60277-4
Abstract

NLRP3 is a pattern recognition receptor forming an inflammasome in response to diverse pathogen and self-derived triggers, but molecular insights on NLRP3 activation are still lacking. Here, we drive ectopic NLRP3 to different subcellular locations in NLRP3-deficient macrophages to map the spatial activation profile of NLRP3, and find that NLRP3 variants enriched at the organellar membranes respond to canonical triggers similarly to wild-type NLRP3; however, unlike wild-type, these NLRP3 variants can be activated even in the absence of the polybasic phospholipid-binding segment. Mechanistically, membrane or protein scaffolds mediate NLRP3 clustering, which leads to the unfastening of the inactive NACHT domain conformation preceding the activated NLRP3 oligomer formation. Our data thus suggest that scaffold-promoted clustering is an important step in NLRP3 activation, enabling NLRP3 to sense distinct activator-induced cellular anomalies exhibited via lipid or protein assemblies, thereby establishing NLRP3 as the master sensor of perturbations in cell homeostasis.

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