Multiple Binding Sites on the Pyrin Domain of ASC Protein Allow Self-association and Interaction with NLRP3 Protein

Vajjhala, Parimala R.; Mirams, Ruth E.; Hill, Jennifer

doi:10.1074/jbc.m112.381228

Cited by 150 publications

(176 citation statements)

References 64 publications

(81 reference statements)

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“…In general terms, this finding agrees with previous mutational studies on ASC PYD oligomerization (16,41). However, several mutations of solvent-exposed residues that abolished ASC PYD self-association do not show, according to our data, the largest variations in chemical shift upon interaction (like Glu-13, Lys-21, and Asp-48, Fig.…”

Section: Asc Pyd Self-associates Through Two Opposing Bindingsupporting

confidence: 82%

“…Electrostatic interactions have been traditionally considered as carrying most of the weight with PYD⅐PYD complex formation (16,18,37,41). As a result, information on the importance of apolar or hydrophobic interactions is scarce.…”

Section: Asc Pyd Self-associates Through Two Opposing Bindingmentioning

confidence: 99%

“…This discrepancy could be explained if the absence of the interaction results from an overall destabilization of the protein structure caused by the mutations, rather than from a perturbation of the interaction interface in the complex. Moreover, in one mutational study, the Thr-63A mutant does not seem to significantly affect ASC selfassociation (16). However, given that Thr-63 is located at the boundaries of the H1, H4, and N-terminal H5 interface (Fig.…”

Section: Asc Pyd Self-associates Through Two Opposing Bindingmentioning

confidence: 99%

“…Despite the abundant structural information reported for PYD domains (the three-dimensional structures of 10 different PYD domains have been deposited in the PDB) (6,17,29,(32)(33)(34)(35)(36)(37)(38), our understanding on the ASC⅐NLRP3 PYD homotypic interaction is very limited. Several models for this interaction have been proposed based on the NLRP3 crystallographic structure and mutational data (4,16), in addition to the recent semi-atomic structure characterization of ASC PYD mature fibril assemblies by cryo-EM and solid-state NMR (18,39).…”

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confidence: 99%

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ASC Pyrin Domain Self-associates and Binds NLRP3 Protein Using Equivalent Binding Interfaces

Oroz

Barrera-Vilarmau

Alfonso³

et al. 2016

Journal of Biological Chemistry

100

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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 ASC⅐ASC and ASC⅐NLRP3 PYD early supramolecular complexes, providing new insights into the molecular mechanisms of inflammasome and apoptosis signaling.Proteins of the death domain superfamily mediate in apoptotic, innate immunity, and inflammatory responses mainly through homotypic interactions (1). Specifically, they are responsible for the assembly of large signaling complexes in which kinases and caspases are activated (1-4). Despite the lack of sequence convergence within the superfamily, its members are characterized by adopting the so-called death fold, consisting of a globular fold with six ␣-helices arranged in a greek-key topology (1, 3). However, some structural differences between the subfamilies are observed within this common fold, including different charge distributions throughout the protein surface, diverse locations of hydrophobic patches, and the different lengths of certain ␣-helices. These differences are thought to lead to homotypic binding specificity (5), albeit certain heterotypic interactions are also known with proposed inhibitory roles in apoptotic and inflammatory pathways (1, 6).Detailed structural information on complexes formed by several members of the death domain superfamily is available. Some examples are the death domain interactions in the PIDDosome (7), the MyDDosome (8), the Fas⅐FADD (9, 10), and the Pelle⅐Tube death domain complexes (11), among others...

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Section: Asc Pyd Self-associates Through Two Opposing Bindingsupporting

confidence: 82%

Section: Asc Pyd Self-associates Through Two Opposing Bindingmentioning

confidence: 99%

Section: Asc Pyd Self-associates Through Two Opposing Bindingmentioning

confidence: 99%

mentioning

confidence: 99%

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ASC Pyrin Domain Self-associates and Binds NLRP3 Protein Using Equivalent Binding Interfaces

Oroz

Barrera-Vilarmau

Alfonso³

et al. 2016

Journal of Biological Chemistry

100

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“…First, it allows formation of an ordered array of IFI16 PYD oligomers on foreign DNA even with the low basal concentration of IFI16 in vivo (38). The IFI16 PYD cluster would then be instrumental for subsequent steps in IFI16 signaling pathways, because assembling PYD/ CARD oligomers by receptor molecules is required for recruiting downstream partners (23,24,39 (28,30). However, we find that extending the size of filaments by conjoining two or more individual oligomers in an end-to-end manner is unique to IFI16 (Figs.…”

Section: Discussionmentioning

confidence: 99%

Cooperative assembly of IFI16 filaments on dsDNA provides insights into host defense strategy

Morrone

Wang²,

Constantoulakis³

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

145

204

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Whether host DNA receptors have any capacity to distinguish self from nonself at the molecular level is an outstanding question in the innate immunity of mammals. Here, by using quantitative assays and electron microscopy, we show that cooperatively assembling into filaments on dsDNA may serve as an integral mechanism by which human IFN-inducible protein-16 (IFI16) engages foreign DNA. IFI16 is essential for defense against a number of different pathogens, and its aberrant activity is also implicated in several autoimmune disorders, such as Sjögren syndrome. IFI16 cooperatively binds dsDNA in a length-dependent manner and clusters into distinct protein filaments even in the presence of excess dsDNA. Consequently, the assembled IFI16·dsDNA oligomers are clearly different from the conventional noninteracting entities resembling beads on a string. The isolated DNA-binding domains of IFI16 engage dsDNA without forming filaments and with weak affinity, and it is the non-DNA-binding pyrin domain of IFI16 that drives the cooperative filament assembly. The surface residues on the pyrin domain that mediate the cooperative DNA binding are conserved, suggesting that related receptors use a common mechanism. These results suggest that IFI16 clusters into signaling foci in a switch-like manner and that it is capable of using the size of naked dsDNA as a molecular ruler to distinguish self from nonself.cooperative filament formation | inflammasome R ecognition of foreign intracellular DNA is a widely conserved defense mechanism by which the innate immune system of mammals detects and responds to invading pathogens (1, 2). Using such a universal molecule as a major danger signal for detecting pathogens must be regulated in a stringent yet efficient manner. However, only a few deciding factors are known for the host innate immune system to selectively engage foreign DNA (i.e., nonself-DNA) while minimizing interactions with self-DNA, which include the compartmentalization of mammalian cells and the size of foreign DNA. These features, however, only raise more questions than provide answers. For example, the footprints of intracellular DNA receptors usually fall below 20 bp, and yet a long foreign DNA fragment [e.g., poly(dA:dT); ≥1,000 bp] is required to induce a robust innate immune response even in a normally DNA-free environment like the cytoplasm (1, 2). On the other hand, foreign DNA-sensing pathways also exist in the host nucleus in which DNA receptors must not respond to abundant self-DNA to prevent spurious activities (1, 2). Indeed, one of the major unresolved questions in understanding the DNA-sensing pathways of mammals is whether the host intracellular DNA receptors have any capacity to distinguish self-from nonself-DNA at the molecular level (2).Human IFN inducible protein-16 (IFI16) is an intracellular DNA receptor of innate immunity that belongs to the family of absent-in-melanoma-2 (AIM2)-like receptors (ALRs) (1-4). IFI16 senses DNA from invading pathogens in both the nucleus and cytoplasm [e.g., vaccinia virus...

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Effects of intestinal lymphatic ligation on intestinal immunity in rats with severe acute pancreatitis

et al. 2021

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Severe acute pancreatitis (SAP) is one of the most common diseases of the gastrointestinal tract, characterized by a complicated pathogenesis, multiple organ failure and high mortality. The primary aim of the present study was to observe the effect of intestinal lymphatic ligation on intestinal injury and modification in rats with severe acute pancreatitis (SAP). Male Sprague-Dawley (SD) rats were randomly divided into: (i) Saline group (SO); (ii) SAP group; and (iii) SAP + ligation group. We evaluated the effect of mesenteric lymphatic duct ligation on the pancreas and intestine tissue by HE. The histopathology of the pancreas in SAP + ligation rats was alleviated slightly compared with SAP rats, but aggravated in the intestine of SAP + ligation rats. Treatment of mesenteric lymphatic duct ligation resulted in an increase in the levels of tumor necrosis factor (TNF)-α, interleukin (IL)-1 and myeloperoxidase (MPO) compared with the small intestinal tissues of SAP rats. In addition, the expression of nucleotide-binding oligomerization domain-like receptors 3 (NLRP3), apoptosis-associated speck-like protein containing a caspase recruitment domain (CARD) domain (ASC) and Caspase-1 in the intestine were higher in the SAP + ligation group. The ratio of Th1/Th2 and regulatory T cells (Tregs) in the mesenteric lymph nodes (MLNs) of the SAP group was lower than those in the SAP + ligation group. The present

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Multiple Binding Sites on the Pyrin Domain of ASC Protein Allow Self-association and Interaction with NLRP3 Protein

Cited by 150 publications

References 64 publications

ASC Pyrin Domain Self-associates and Binds NLRP3 Protein Using Equivalent Binding Interfaces

ASC Pyrin Domain Self-associates and Binds NLRP3 Protein Using Equivalent Binding Interfaces

Cooperative assembly of IFI16 filaments on dsDNA provides insights into host defense strategy

Effects of intestinal lymphatic ligation on intestinal immunity in rats with severe acute pancreatitis

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