PDBsum entry 2n1f

Apoptosis

PDB id: 2n1f

Contents

Protein chains

(+ 9 more) 89 a.a.

PDB id:

2n1f

Name:

Apoptosis

Title:

Structure and assembly of the mouse asc filament by combined nmr spectroscopy and cryo-electron microscopy

Structure:

Apoptosis-associated speck-like protein. Chain: a, b, c, d, e, f, g, h, i, j, k, l, m, n, o. Fragment: pyrin domain (unp residues 2-90). Synonym: masc. Engineered: yes

Source:

Mus musculus. Mouse. Organism_taxid: 10090. Gene: pycard, asc. Expressed in: escherichia coli bl21(de3). Expression_system_taxid: 469008.

NMR struc:

10 models

Authors:

L.Sborgi,F.Ravotti,V.Dandey,M.Dick,A.Mazur,S.Reckel,M.Chami, S.Scherer,A.Bockmann,E.Egelman,H.Stahlberg,P.Broz,B.Meier,S.Hiller

Key ref:

L.Sborgi et al. (2015). Structure and assembly of the mouse ASC inflammasome by combined NMR spectroscopy and cryo-electron microscopy. Proc Natl Acad Sci U S A, 112, 13237-13242. PubMed id: 26464513 DOI: 10.1073/pnas.1507579112

Date:

01-Apr-15 Release date: 14-Oct-15

Protein chains

Q9EPB4  (ASC_MOUSE) -  Apoptosis-associated speck-like protein containing a CARD from Mus musculus

Seq: 193 a.a.

Struc: 89 a.a.

Enzyme reactions

• Enzyme class: E.C.?

DOI no: 10.1073/pnas.1507579112

Proc Natl Acad Sci U S A 112:13237-13242 (2015)

PubMed id: 26464513

Structure and assembly of the mouse ASC inflammasome by combined NMR spectroscopy and cryo-electron microscopy.

L.Sborgi, F.Ravotti, V.P.Dandey, M.S.Dick, A.Mazur, S.Reckel, M.Chami, S.Scherer, M.Huber, A.Böckmann, E.H.Egelman, H.Stahlberg, P.Broz, B.H.Meier, S.Hiller.

ABSTRACT

Inflammasomes are multiprotein complexes that control the innate immune response by activating caspase-1, thus promoting the secretion of cytokines in response to invading pathogens and endogenous triggers. Assembly of inflammasomes is induced by activation of a receptor protein. Many inflammasome receptors require the adapter protein ASC [apoptosis-associated speck-like protein containing a caspase-recruitment domain (CARD)], which consists of two domains, the N-terminal pyrin domain (PYD) and the C-terminal CARD. Upon activation, ASC forms large oligomeric filaments, which facilitate procaspase-1 recruitment. Here, we characterize the structure and filament formation of mouse ASC in vitro at atomic resolution. Information from cryo-electron microscopy and solid-state NMR spectroscopy is combined in a single structure calculation to obtain the atomic-resolution structure of the ASC filament. Perturbations of NMR resonances upon filament formation monitor the specific binding interfaces of ASC-PYD association. Importantly, NMR experiments show the rigidity of the PYD forming the core of the filament as well as the high mobility of the CARD relative to this core. The findings are validated by structure-based mutagenesis experiments in cultured macrophages. The 3D structure of the mouse ASC-PYD filament is highly similar to the recently determined human ASC-PYD filament, suggesting evolutionary conservation of ASC-dependent inflammasome mechanisms.