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@ARTICLE{Oroz:138760,
author = {Oroz, Javier and Barrera-Vilarmau, Susana and Alfonso,
Carlos and Rivas, Germán and de Alba, Eva},
title = {{ASC} {P}yrin {D}omain {S}elf-associates and {B}inds
{NLRP}3 {P}rotein {U}sing {E}quivalent {B}inding
{I}nterfaces.},
journal = {The journal of biological chemistry},
volume = {291},
number = {37},
issn = {0021-9258},
address = {Bethesda, Md.},
publisher = {Soc.60645},
reportid = {DZNE-2020-05082},
pages = {19487-19501},
year = {2016},
abstract = {Death domain superfamily members typically act as adaptors
mediating in the assembly of supramolecular complexes with
critical apoptosis and inflammation functions. These modular
proteins consist of death domains, death effector domains,
caspase recruitment domains, and pyrin domains (PYD).
Despite the high structural similarity among them, only
homotypic interactions participate in complex formation,
suggesting that subtle factors differentiate each
interaction type. It is thus critical to identify these
factors as an essential step toward the understanding of the
molecular basis of apoptosis and inflammation. The proteins
apoptosis-associated speck-like protein containing a CARD
(ASC) and NLRP3 play key roles in the regulation of
apoptosis and inflammation through self-association and
protein-protein interactions mediated by their PYDs. To
better understand the molecular basis of their function, we
have characterized ASC and NLRP3 PYD self-association and
their intermolecular interaction by solution NMR
spectroscopy and analytical ultracentrifugation. We found
that ASC self-associates and binds NLRP3 PYD through
equivalent protein regions, with higher binding affinity for
the latter. These regions are located at opposite sides of
the protein allowing multimeric complex formation previously
shown in ASC PYD fibril assemblies. We show that NLRP3 PYD
coexists in solution as a monomer and highly populated
large-order oligomerized species. Despite this, we
determined its monomeric three-dimensional solution
structure by NMR and characterized its binding to ASC PYD.
Using our novel structural data, we propose molecular models
of ASC·ASC and ASC·NLRP3 PYD early supramolecular
complexes, providing new insights into the molecular
mechanisms of inflammasome and apoptosis signaling.},
keywords = {CARD Signaling Adaptor Proteins / Cytoskeletal Proteins:
chemistry / Cytoskeletal Proteins: genetics / Cytoskeletal
Proteins: metabolism / Humans / Models, Molecular / NLR
Family, Pyrin Domain-Containing 3 Protein: chemistry / NLR
Family, Pyrin Domain-Containing 3 Protein: genetics / NLR
Family, Pyrin Domain-Containing 3 Protein: metabolism /
Nuclear Magnetic Resonance, Biomolecular / Protein Binding /
Protein Structure, Quaternary / Pyrin Domain /
Ultracentrifugation / CARD Signaling Adaptor Proteins (NLM
Chemicals) / Cytoskeletal Proteins (NLM Chemicals) / NLR
Family, Pyrin Domain-Containing 3 Protein (NLM Chemicals) /
NLRP3 protein, human (NLM Chemicals) / PYCARD protein, human
(NLM Chemicals)},
cin = {AG Zweckstetter},
ddc = {540},
cid = {I:(DE-2719)1410001},
pnm = {342 - Disease Mechanisms and Model Systems (POF3-342)},
pid = {G:(DE-HGF)POF3-342},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:27432880},
pmc = {pmc:PMC5016686},
doi = {10.1074/jbc.M116.741082},
url = {https://pub.dzne.de/record/138760},
}