Pyroptosis inhibiting nanobodies block Gasdermin D pore formation.
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
01 Dec 2023
01 Dec 2023
Historique:
received:
18
04
2023
accepted:
16
11
2023
medline:
4
12
2023
pubmed:
2
12
2023
entrez:
1
12
2023
Statut:
epublish
Résumé
Human Gasdermin D (GSDMD) is a key mediator of pyroptosis, a pro-inflammatory form of cell death occurring downstream of inflammasome activation as part of the innate immune defence. Upon cleavage by inflammatory caspases in the cytosol, the N-terminal domain of GSDMD forms pores in the plasma membrane resulting in cytokine release and eventually cell death. Targeting GSDMD is an attractive way to dampen inflammation. In this study, six GSDMD targeting nanobodies are characterized in terms of their binding affinity, stability, and effect on GSDMD pore formation. Three of the nanobodies inhibit GSDMD pore formation in a liposome leakage assay, although caspase cleavage was not perturbed. We determine the crystal structure of human GSDMD in complex with two nanobodies at 1.9 Å resolution, providing detailed insights into the GSDMD-nanobody interactions and epitope binding. The pore formation is sterically blocked by one of the nanobodies that binds to the oligomerization interface of the N-terminal domain in the multi-subunit pore assembly. Our biochemical and structural findings provide tools for studying inflammasome biology and build a framework for the design of GSDMD targeting drugs.
Identifiants
pubmed: 38040708
doi: 10.1038/s41467-023-43707-z
pii: 10.1038/s41467-023-43707-z
pmc: PMC10692205
doi:
Substances chimiques
Caspases
EC 3.4.22.-
Gasdermins
0
Inflammasomes
0
Intracellular Signaling Peptides and Proteins
0
Phosphate-Binding Proteins
0
Single-Domain Antibodies
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
7923Subventions
Organisme : Deutsche Forschungsgemeinschaft (German Research Foundation)
ID : EXC2151-390873048
Organisme : Deutsche Forschungsgemeinschaft (German Research Foundation)
ID : GRK 2168
Informations de copyright
© 2023. The Author(s).
Références
Science. 2020 Sep 25;369(6511):1633-1637
pubmed: 32820063
Nature. 2023 Jun;618(7967):1072-1077
pubmed: 37196676
J Mol Biol. 2022 Feb 28;434(4):167246
pubmed: 34537232
Nat Commun. 2023 Jan 4;14(1):61
pubmed: 36599845
Acta Crystallogr D Biol Crystallogr. 2004 Dec;60(Pt 12 Pt 1):2126-32
pubmed: 15572765
Nat Rev Drug Discov. 2021 May;20(5):384-405
pubmed: 33692549
Nucleic Acids Res. 2014 Jul;42(Web Server issue):W320-4
pubmed: 24753421
Immunity. 2019 Jul 16;51(1):43-49.e4
pubmed: 31097341
Science. 2018 Nov 23;362(6417):956-960
pubmed: 30467171
Nature. 1993 Jun 3;363(6428):446-8
pubmed: 8502296
Nat Rev Immunol. 2020 Mar;20(3):143-157
pubmed: 31690840
Nat Protoc. 2014 Mar;9(3):674-93
pubmed: 24577359
Pharmaceuticals (Basel). 2021 Jun 23;14(7):
pubmed: 34201507
Front Immunol. 2021 Nov 11;12:751533
pubmed: 34858408
Annu Rev Immunol. 2018 Apr 26;36:695-715
pubmed: 29490163
Cell Host Microbe. 2021 Oct 13;29(10):1521-1530.e10
pubmed: 34492225
J Exp Med. 2016 May 2;213(5):771-90
pubmed: 27069117
Acta Crystallogr D Biol Crystallogr. 2010 Jan;66(Pt 1):12-21
pubmed: 20057044
Nature. 2023 Apr;616(7957):598-605
pubmed: 36991125
Eur J Immunol. 2018 Apr;48(4):584-592
pubmed: 29274245
Cell Rep. 2020 Mar 17;30(11):3743-3754.e6
pubmed: 32187546
Nat Chem. 2017 Aug;9(8):762-771
pubmed: 28754949
Nature. 2021 May;593(7860):607-611
pubmed: 33883744
Semin Immunol. 2023 Sep;69:101811
pubmed: 37473560
Neurobiol Dis. 2020 Feb;134:104619
pubmed: 31669671
Mol Ther Methods Clin Dev. 2020 Jan 30;17:401-408
pubmed: 32128345
Nature. 2016 Jul 06;535(7610):153-8
pubmed: 27383986
Science. 2020 May 29;368(6494):
pubmed: 32299851
Cell. 2021 Aug 19;184(17):4495-4511.e19
pubmed: 34289345
Ther Adv Neurol Disord. 2015 Jan;8(1):20-30
pubmed: 25584071
Chem Res Toxicol. 1997 Dec;10(12):1319-24
pubmed: 9437520
J Immunol. 2019 Aug 1;203(3):736-748
pubmed: 31209100
Immunity. 2018 Jan 16;48(1):35-44.e6
pubmed: 29195811
Nat Commun. 2022 Jan 10;13(1):52
pubmed: 35013201
J Mol Biol. 2007 Sep 21;372(3):774-97
pubmed: 17681537
Free Radic Biol Med. 2020 May 20;152:8-17
pubmed: 32151746
Front Immunol. 2021 Oct 14;12:750841
pubmed: 34721422
Protein Sci. 2002 Mar;11(3):500-15
pubmed: 11847273
Acta Crystallogr D Biol Crystallogr. 2010 Feb;66(Pt 2):213-21
pubmed: 20124702
Nature. 2015 Oct 29;526(7575):666-71
pubmed: 26375259
Acta Crystallogr D Biol Crystallogr. 2010 Feb;66(Pt 2):125-32
pubmed: 20124692
Mol Cell. 2002 Aug;10(2):417-26
pubmed: 12191486
Elife. 2022 Nov 14;11:
pubmed: 36374182
Front Cell Dev Biol. 2021 Jul 28;9:715211
pubmed: 34395445
Nature. 2021 Mar;591(7848):131-136
pubmed: 33472215
Nature. 2015 Oct 29;526(7575):660-5
pubmed: 26375003
Nature. 2011 Oct 16;479(7371):117-21
pubmed: 22002608
Sci Rep. 2016 Jan 21;6:19211
pubmed: 26791954
EMBO J. 2016 Aug 15;35(16):1766-78
pubmed: 27418190
Nature. 2016 Jul 7;535(7610):111-6
pubmed: 27281216
Nature. 2023 Jun;618(7967):1065-1071
pubmed: 37198476
Nat Commun. 2017 Jan 03;8:14128
pubmed: 28045099
Brain Behav. 2021 Apr;11(4):e02063
pubmed: 33587329
Nat Cell Biol. 2020 Oct;22(10):1264-1275
pubmed: 32929201
Sci Immunol. 2018 Aug 24;3(26):
pubmed: 30143556
Sci Rep. 2019 Dec 10;9(1):18688
pubmed: 31822703
Nat Immunol. 2020 Jul;21(7):736-745
pubmed: 32367036