IntAct European Bioinformatics Institute IntAct intact http://www.ebi.ac.uk/ http://www.ebi.ac.uk/intact/query/${ac} EBI-[0-9]+|IA:[0-9]+ INTerAction database (IntAct) provides an open source database and toolkit for the storage, presentation and analysis of molecular interactions. http://www.ebi.ac.uk/intact European Bioinformatics Institute; Wellcome Trust Genome Campus; Hinxton, Cambridge; CB10 1SD; United Kingdom http://www.ebi.ac.uk/intact/ http://www.ebi.ac.uk/ lu-2014-4 Unified polymerization mechanism for the assembly of ASC-dependent inflammasomes. in vitro In vitro fps fluorescence polarization spectroscopy Fluorescence anisotropy FPS FPS Fluorescence anisotropy fps tag fluorescence tag visualisation by fluorescence tag fluorescence Lu A., Magupalli V.G., Ruan J., Yin Q., Atianand M.K., Vos M.R., Schröder G.F., Fitzgerald K.A., Wu H., Egelman E.H. A His-MBP-ASCPYD fusion construct with an added C-terminal Cys for conjugating with Alexa 488 fluorophore was purified. The large fusion tag MBP inhibited ASC-PYD polymerization to enable His-MBP-ASC-PYD to be expressed in the monomeric form. Polymerization of Alexa-488-labeled monomeric ASCPYD was initiated by addition of the TEV protease to remove His-MBP from the fusion protein. Origin of AIM2, NRPL3 and Caspase-1 taken from sequences given in Supp. Fig. 3D and from previous work by same author (PMID:191586750) hao.wu@childrens.harvard.edu Cell (0092-8674) 2014 Only protein-protein interactions imex curation lu-2014-5 Unified polymerization mechanism for the assembly of ASC-dependent inflammasomes. in vitro In vitro electron microscopy electron microscopy Electron crystallography Electron cryomicroscopy Electron crystallography Electron cryomicroscopy predetermined predetermined participant predetermined Lu A., Magupalli V.G., Ruan J., Yin Q., Atianand M.K., Vos M.R., Schröder G.F., Fitzgerald K.A., Wu H., Egelman E.H. The complex was purified by Ni-NTA resin followed by gel filtration. Origin of AIM2, NRPL3 and Caspase-1 taken from sequences given in Supp. Fig. 3D and from previous work by same author (PMID:191586750) hao.wu@childrens.harvard.edu Cell (0092-8674) 2014 Only protein-protein interactions imex curation lu-2014-2 Unified polymerization mechanism for the assembly of ASC-dependent inflammasomes. ecoli Escherichia coli (strain K12) molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column weight by comassie molecular weight estimation by coomasie staining weight by comassie Lu A., Magupalli V.G., Ruan J., Yin Q., Atianand M.K., Vos M.R., Schröder G.F., Fitzgerald K.A., Wu H., Egelman E.H. The complex was purified by Ni-NTA resin followed by gel filtration. Origin of AIM2, NRPL3 and Caspase-1 taken from sequences given in Supp. Fig. 3D and from previous work by same author (PMID:191586750) hao.wu@childrens.harvard.edu Cell (0092-8674) 2014 Only protein-protein interactions imex curation lu-2014-7 Unified polymerization mechanism for the assembly of ASC-dependent inflammasomes. in vitro In vitro pull down pull down weight by comassie molecular weight estimation by coomasie staining weight by comassie Lu A., Magupalli V.G., Ruan J., Yin Q., Atianand M.K., Vos M.R., Schröder G.F., Fitzgerald K.A., Wu H., Egelman E.H. Origin of AIM2, NRPL3 and Caspase-1 taken from sequences given in Supp. Fig. 3D and from previous work by same author (PMID:191586750) hao.wu@childrens.harvard.edu Cell (0092-8674) 2014 Only protein-protein interactions imex curation lu-2014-10 in vitro In vitro electron microscopy electron microscopy Electron crystallography Electron cryomicroscopy Electron crystallography Electron cryomicroscopy tag visualisation tag visualisation Lu A., Magupalli V.G., Ruan J., Yin Q., Atianand M.K., Vos M.R., Schröder G.F., Fitzgerald K.A., Wu H., Egelman E.H. Origin of AIM2, NRPL3 and Caspase-1 taken from sequences given in Supp. Fig. 3D and from previous work by same author (PMID:191586750) hao.wu@childrens.harvard.edu Cell (0092-8674) 2014 Only protein-protein interactions imex curation lu-2014-1 Unified polymerization mechanism for the assembly of ASC-dependent inflammasomes. ecoli Escherichia coli (strain K12) electron microscopy electron microscopy Electron crystallography Electron cryomicroscopy Electron crystallography Electron cryomicroscopy predetermined predetermined participant predetermined Lu A., Magupalli V.G., Ruan J., Yin Q., Atianand M.K., Vos M.R., Schröder G.F., Fitzgerald K.A., Wu H., Egelman E.H. The complex was purified by Ni-NTA resin followed by gel filtration. Origin of AIM2, NRPL3 and Caspase-1 taken from sequences given in Supp. Fig. 3D and from previous work by same author (PMID:191586750) hao.wu@childrens.harvard.edu Cell (0092-8674) 2014 Only protein-protein interactions imex curation lu-2014-3 Unified polymerization mechanism for the assembly of ASC-dependent inflammasomes. ecoli Escherichia coli (strain K12) electron microscopy electron microscopy Electron crystallography Electron cryomicroscopy Electron crystallography Electron cryomicroscopy immunostaining immunostaining Lu A., Magupalli V.G., Ruan J., Yin Q., Atianand M.K., Vos M.R., Schröder G.F., Fitzgerald K.A., Wu H., Egelman E.H. The complex was purified by Ni-NTA resin followed by gel filtration. Origin of AIM2, NRPL3 and Caspase-1 taken from sequences given in Supp. Fig. 3D and from previous work by same author (PMID:191586750) hao.wu@childrens.harvard.edu Cell (0092-8674) 2014 Only protein-protein interactions imex curation lu-2014-6 Unified polymerization mechanism for the assembly of ASC-dependent inflammasomes. in vitro In vitro electron microscopy electron microscopy Electron crystallography Electron cryomicroscopy Electron crystallography Electron cryomicroscopy immunostaining immunostaining Lu A., Magupalli V.G., Ruan J., Yin Q., Atianand M.K., Vos M.R., Schröder G.F., Fitzgerald K.A., Wu H., Egelman E.H. Origin of AIM2, NRPL3 and Caspase-1 taken from sequences given in Supp. Fig. 3D and from previous work by same author (PMID:191586750) hao.wu@childrens.harvard.edu Cell (0092-8674) 2014 Only protein-protein interactions imex curation lu-2014-8 Unified polymerization mechanism for the assembly of ASC-dependent inflammasomes. human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Lu A., Magupalli V.G., Ruan J., Yin Q., Atianand M.K., Vos M.R., Schröder G.F., Fitzgerald K.A., Wu H., Egelman E.H. A His-MBP-ASCPYD fusion construct with an added C-terminal Cys for conjugating with Alexa 488 fluorophore was purified. The large fusion tag MBP inhibited ASC-PYD polymerization to enable His-MBP-ASC-PYD to be expressed in the monomeric form. Polymerization of Alexa-488-labeled monomeric ASCPYD was initiated by addition of the TEV protease to remove His-MBP from the fusion protein. Origin of AIM2, NRPL3 and Caspase-1 taken from sequences given in Supp. Fig. 3D and from previous work by same author (PMID:191586750) hao.wu@childrens.harvard.edu Cell (0092-8674) 2014 Only protein-protein interactions imex curation lu-2014-9 Unified polymerization mechanism for the assembly of ASC-dependent inflammasomes. human-thp-1 Homo sapiens acute monocytic leukemia cells thp-1 Human acute monocytic leukemia cell line Human acute monocytic leukemia established from the peripheral blood of a 1-year-old boy with acute monocytic leukemia (AML) at relapse in 1978. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Lu A., Magupalli V.G., Ruan J., Yin Q., Atianand M.K., Vos M.R., Schröder G.F., Fitzgerald K.A., Wu H., Egelman E.H. Cells treated with uric-acid crystals Origin of AIM2, NRPL3 and Caspase-1 taken from sequences given in Supp. Fig. 3D and from previous work by same author (PMID:191586750) hao.wu@childrens.harvard.edu Cell (0092-8674) 2014 Only protein-protein interactions imex curation asc_human Apoptosis-associated speck-like protein containing a CARD PYD and CARD domain-containing protein Target of methylation-induced silencing 1 Caspase recruitment domain-containing protein 5 ASC CARD5 TMS1 PYCARD protein protein human Homo sapiens Human MGRARDAILDALENLTAEELKKFKLKLLSVPLREGYGRIPRGALLSMDALDLTDKLVSFYLETYGAELTANVLRDMGLQEMAGQLQAATHQGSGAAPAGIQAPPQSAAKPGLHFIDQHRAALIARVTNVEWLLDALYGKVLTDEQYQAVRAEPTNPSKMRKLFSFTPAWNWTCKDLLLQALRESQSYLVEDLERS aim2_human Interferon-inducible protein AIM2 AIM2 Absent in melanoma 2 protein protein human Homo sapiens Human MESKYKEILLLTGLDNITDEELDRFKFFLSDEFNIATGKLHTANRIQVATLMIQNAGAVSAVMKTIRIFQKLNYMLLAKRLQEEKEKVDKQYKSVTKPKPLSQAEMSPAASAAIRNDVAKQRAAPKVSPHVKPEQKQMVAQQESIREGFQKRCLPVMVLKAKKPFTFETQEGKQEMFHATVATEKEFFFVKVFNTLLKDKFIPKRIIIIARYYRHSGFLEVNSASRVLDAESDQKVNVPLNIIRKAGETPKINTLQTQPLGTIVNGLFVVQKVTEKKKNILFDLSDNTGKMEVLGVRNEDTMKCKEGDKVRLTFFTLSKNGEKLQLTSGVHSTIKVIKAKKKT nlrp3_human NACHT, LRR and PYD domains-containing protein 3 NLRP3 C1orf7 CIAS1 NALP3 PYPAF1 Angiotensin/vasopressin receptor AII/AVP-like Caterpiller protein 1.1 Cryopyrin PYRIN-containing APAF1-like protein 1 Cold-induced autoinflammatory syndrome 1 protein protein protein human Homo sapiens Human MKMASTRCKLARYLEDLEDVDLKKFKMHLEDYPPQKGCIPLPRGQTEKADHVDLATLMIDFNGEEKAWAMAVWIFAAINRRDLYEKAKRDEPKWGSDNARVSNPTVICQEDSIEEEWMGLLEYLSRISICKMKKDYRKKYRKYVRSRFQCIEDRNARLGESVSLNKRYTRLRLIKEHRSQQEREQELLAIGKTKTCESPVSPIKMELLFDPDDEHSEPVHTVVFQGAAGIGKTILARKMMLDWASGTLYQDRFDYLFYIHCREVSLVTQRSLGDLIMSCCPDPNPPIHKIVRKPSRILFLMDGFDELQGAFDEHIGPLCTDWQKAERGDILLSSLIRKKLLPEASLLITTRPVALEKLQHLLDHPRHVEILGFSEAKRKEYFFKYFSDEAQARAAFSLIQENEVLFTMCFIPLVCWIVCTGLKQQMESGKSLAQTSKTTTAVYVFFLSSLLQPRGGSQEHGLCAHLWGLCSLAADGIWNQKILFEESDLRNHGLQKADVSAFLRMNLFQKEVDCEKFYSFIHMTFQEFFAAMYYLLEEEKEGRTNVPGSRLKLPSRDVTVLLENYGKFEKGYLIFVVRFLFGLVNQERTSYLEKKLSCKISQQIRLELLKWIEVKAKAKKLQIQPSQLELFYCLYEMQEEDFVQRAMDYFPKIEINLSTRMDHMVSSFCIENCHRVESLSLGFLHNMPKEEEEEEKEGRHLDMVQCVLPSSSHAACSHGLVNSHLTSSFCRGLFSVLSTSQSLTELDLSDNSLGDPGMRVLCETLQHPGCNIRRLWLGRCGLSHECCFDISLVLSSNQKLVELDLSDNALGDFGIRLLCVGLKHLLCNLKKLWLVSCCLTSACCQDLASVLSTSHSLTRLYVGENALGDSGVAILCEKAKNPQCNLQKLGLVNSGLTSVCCSALSSVLSTNQNLTHLYLRGNTLGDKGIKLLCEGLLHPDCKLQVLELDNCNLTSHCCWDLSTLLTSSQSLRKLSLGNNDLGDLGVMMFCEVLKQQSCLLQNLGLSEMYFNYETKSALETLQEEKPELTVVFEPSW casp1_human Caspase-1 Interleukin-1 beta convertase Interleukin-1 beta-converting enzyme p45 CASP1 IL1BC IL1BCE protein protein human Homo sapiens Human MADKVLKEKRKLFIRSMGEGTINGLLDELLQTRVLNKEEMEKVKRENATVMDKTRALIDSVIPKGAQACQICITYICEEDSYLAGTLGLSADQTSGNYLNMQDSQGVLSSFPAPQAVQDNPAMPTSSGSEGNVKLCSLEEAQRIWKQKSAEIYPIMDKSSRTRLALIICNEEFDSIPRRTGAEVDITGMTMLLQNLGYSVDVKKNLTASDMTTELEAFAHRPEHKTSDSTFLVFMSHGIREGICGKKHSEQVPDILQLNAIFNMLNTKNCPSLKDKPKVIIIQACRGDSPGVVWFKDSVGVSGNLSLPTTEEFEDDAIKKAHIEKDFIAFCSSTPDNVSWRHPTMGSVFIGRLIEHMQEYACSCDVEEIFRKVRFSFEQPDGRAQMPTTERVTLTRCFYLFPGH pycard-pycard-4 1537366 n/a 1537376 unspecified role unspecified role neutral component neutral component pyd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false c-terminal alexa 488 label alexa 488 label c-term range C-terminal range c-term range c-term range C-terminal range c-term range false ecoli Escherichia coli (strain K12) n/a 1537376 unspecified role unspecified role neutral component neutral component pyd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false c-terminal alexa 488 label alexa 488 label c-term range C-terminal range c-term range c-term range C-terminal range c-term range false ecoli Escherichia coli (strain K12) 1537385 1537382 direct interaction direct interaction false false false Fig. 1E, 1F aim2-pycard-5 1537366 n/a 1537377 unspecified role unspecified role neutral component neutral component pyd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false n-terminal mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false O14862:p.Leu10_Leu11delinsAlaAla mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false O14862:p.Gly38Glu mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false O14862:p.Tyr74Arg mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false O14862:p.Phe27Gly mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false O14862:p.Arg24Glu mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false O14862:p.Asp15Arg mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false O14862:p.Lys39Glu mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1537376 unspecified role unspecified role neutral component neutral component pyd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false c-terminal alexa 488 label alexa 488 label c-term range C-terminal range c-term range c-term range C-terminal range c-term range false ecoli Escherichia coli (strain K12) 1537400 1537389 direct interaction direct interaction false false false The rates of polymerization were dramatically enhanced in the presence of increasing amounts of sub-stoichiometric AIM2. Fig. 1E, 5E aim2-pycard-6 1537366 n/a 1537376 unspecified role unspecified role neutral component neutral component pyd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false c-terminal alexa 488 label alexa 488 label c-term range C-terminal range c-term range c-term range C-terminal range c-term range false ecoli Escherichia coli (strain K12) n/a 1537377 unspecified role unspecified role neutral component neutral component n-terminal mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false The rates of polymerization were dramatically enhanced in the presence of increasing amounts of sub-stoichiometric AIM2 Fig. 1F Double-stranded DNA nlrp3-pycard-6 1537366 n/a 1537376 unspecified role unspecified role neutral component neutral component c-terminal alexa 488 label alexa 488 label c-term range C-terminal range c-term range c-term range C-terminal range c-term range false pyd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1537378 unspecified role unspecified role neutral component neutral component pyd-nbd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n-terminal mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false Q96P20:p.Arg43Trp mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false Q96P20:p.Met27Glu mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false Q96P20:p.Glu64Arg mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false Q96P20:p.Asp82Arg mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false Q96P20:p.Glu15Arg mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false Q96P20:p.Lys23_Lys24delinsGluGlu mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) 1537414 1537412 direct interaction direct interaction false false false ASC PYD-domian did not significantly polymerize unless increasing amounts of NLRP3 were added Fig. 2B, 2D, 5F Supp. Fig. 2A casp1-casp1-1 1537366 n/a 1537379 unspecified role unspecified role neutral component neutral component purified purified card domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sumoylated lysine sumoylated lysine undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) n/a 1537379 unspecified role unspecified role neutral component neutral component purified purified card domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sumoylated lysine sumoylated lysine undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) 1537430 1537425 direct interaction direct interaction false false false Fig.6F casp1-pycard-1 1537366 n/a 1537376 unspecified role unspecified role neutral component neutral component purified purified card domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1537379 unspecified role unspecified role neutral component neutral component purified purified region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false card domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region sumoylated lysine sumoylated lysine undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false 1537439 1537436 direct interaction direct interaction false false false In the presence ASC sub-stoichiometric ratios, caspase-1 polymerization was greatly enhanced, consistent with nucleation of caspase-1 polymerization by ASC. Fig.6F nlrp3-pycard-1 1537366 N/A 1537376 unspecified role unspecified role neutral component neutral component c-terminal alexa 488 label alexa 488 label c-term range C-terminal range c-term range c-term range C-terminal range c-term range false pyd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 1537378 unspecified role unspecified role neutral component neutral component n-terminal mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false direct interaction direct interaction false false false Fig. 2D Full-length MLRP3 was expressed in an unspecified insect cell line nlrp3-pycard-3 1537366 N/A 1537376 unspecified role unspecified role neutral component neutral component c-terminal alexa 488 label alexa 488 label c-term range C-terminal range c-term range c-term range C-terminal range c-term range false pyd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 1537378 unspecified role unspecified role neutral component neutral component n-terminal mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false trini-high_5 Trichoplusia ni cell line from eggs high_5 Insect cell line from Trichoplusia ni egg cells Invitrogen supplies this cell line. Invitrogen supply this cell line. direct interaction direct interaction false false false ASC PYD-domian did not significantly polymerize unless increasing amounts of NLRP3 were added Fig. 2C aim2-aim2 1537367 n/a 1537377 unspecified role unspecified role neutral component neutral component purified purified region sumoylated lysine sumoylated lysine undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n-terminal mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false pyd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) n/a 1537377 unspecified role unspecified role neutral component neutral component purified purified n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false pyd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n-terminal mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false region sumoylated lysine sumoylated lysine undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) 1537464 1537460 direct interaction direct interaction false false false Supp Fig. 1B aim2-pycard-7 1537367 n/a 1537376 unspecified role unspecified role neutral component neutral component purified purified pyd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false c-terminal alexa 488 label alexa 488 label c-term range C-terminal range c-term range c-term range C-terminal range c-term range false ecoli Escherichia coli (strain K12) n/a 1537377 unspecified role unspecified role neutral component neutral component purified purified pyd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false n-terminal mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) 1537472 1537469 direct interaction direct interaction false false false Supp. Fig. 1F nlrp3-nlrp3 1537367 n/a 1537378 unspecified role unspecified role neutral component neutral component purified purified pyd-nbd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n-terminal mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) n/a 1537378 unspecified role unspecified role neutral component neutral component purified purified pyd-nbd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false n-terminal mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) 1537481 1537477 direct interaction direct interaction false false false Supp. Fig. 2C pycard-pycard-1 1537367 n/a 1537376 unspecified role unspecified role neutral component neutral component purified purified pyd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false c-terminal alexa 488 label alexa 488 label c-term range C-terminal range c-term range c-term range C-terminal range c-term range false ecoli Escherichia coli (strain K12) n/a 1537376 unspecified role unspecified role neutral component neutral component purified purified pyd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false c-terminal alexa 488 label alexa 488 label c-term range C-terminal range c-term range c-term range C-terminal range c-term range false ecoli Escherichia coli (strain K12) 1537489 1537486 direct interaction direct interaction false false false Fig. 3A, supp. Fig 4I High salt concentrations pycard-pycard-8 1537367 n/a 1537376 unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) n/a 1537376 unspecified role unspecified role neutral component neutral component purified purified ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false Supp. Fig. 6A aim2-pycard-2 1537368 n/a 1537376 unspecified role unspecified role neutral component neutral component n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false pyd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n/a 1537377 anti tag western anti tag western blot anti tag western unspecified role unspecified role neutral component neutral component n-terminal biotin tag biotin tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false pyd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 1537500 1537497 physical association physical association false false false Fig. 1C, Supp 1A The complex eluted at the void position of a Superdex 200 gel filtration column suggesting formation of large aggregates. pycard-pycard-2 1537368 n/a 1537376 unspecified role unspecified role neutral component neutral component Q9ULZ3:p.Lys21Gln mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false Q9ULZ3:p.Asp48Arg mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false Q9ULZ3:p.Phe59Glu mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false Q9ULZ3:p.Lys21_Lys22delinsGluGlu mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false pyd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Q9ULZ3:p.Asp48Asn mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false Q9ULZ3:p.Glu13Arg mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false n-terminal mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false Q9ULZ3:p.Lys26Glu mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false Q9ULZ3:p.Arg41Glu mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false n/a 1537376 unspecified role unspecified role neutral component neutral component Q9ULZ3:p.Asp48Arg mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false Q9ULZ3:p.Asp48Asn mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false Q9ULZ3:p.Glu13Arg mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false Q9ULZ3:p.Lys26Glu mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false n-terminal mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false Q9ULZ3:p.Phe59Glu mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false Q9ULZ3:p.Lys21Gln mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false Q9ULZ3:p.Lys21_Lys22delinsGluGlu mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false pyd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Q9ULZ3:p.Arg41Glu mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false 1537523 1537507 direct interaction direct interaction false false false Fig. 5A casp1-aim2-3 1537369 n/a 1537376 unspecified role unspecified role prey prey purified purified ecoli Escherichia coli (strain K12) n/a 1537379 unspecified role unspecified role bait bait purified purified region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false card domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1537377 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey purified purified pyd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) physical association physical association false false false Fig.6C casp1-aim2-2 1537369 n/a 1537376 unspecified role unspecified role prey prey purified purified ecoli Escherichia coli (strain K12) n/a 1537379 unspecified role unspecified role bait bait purified purified region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false card domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false Supp Fig.6C nlrp3-pycard-7 1537370 n/a 1537378 unspecified role unspecified role neutral component neutral component region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n-terminal mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false pyd-nbd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1537376 unspecified role unspecified role neutral component neutral component purified purified c-terminal alexa 488 label alexa 488 label c-term range C-terminal range c-term range c-term range C-terminal range c-term range false pyd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) 1537548 1537545 direct interaction direct interaction false false false Fig. 2E aim2-pycard-1 1537371 n/a 1537377 unspecified role unspecified role neutral component neutral component biotin acceptor peptide tag tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false pyd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n/a 1537376 unspecified role unspecified role neutral component neutral component n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false pyd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 1537555 1537552 physical association physical association false false false Fig. 1B aim2-pycard-3 1537372 n/a 1537376 unspecified role unspecified role neutral component neutral component pyd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false n/a 1537377 unspecified role unspecified role neutral component neutral component pyd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n-terminal biotin tag biotin tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false 1537561 1537558 physical association physical association false false false Fig. 1D, SUpp. Fig. 1C Detection repeated using both streptavidin-gold conjugate (6 nm) and Ni-NTA-gold (5 nm) casp1-aim2-1 1537373 n/a 1537377 unspecified role unspecified role prey prey purified purified ecoli Escherichia coli (strain K12) n/a 1537379 unspecified role unspecified role bait bait purified purified region gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false card domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) n/a 1537376 unspecified role unspecified role prey prey purified purified pyd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) physical association physical association false false false Fig. 6D, 6E Complex visualised with both anti-ASC gold and Ni-NTA gold pycard-pycard-3 1537374 n/a 1537376 unspecified role unspecified role bait bait n-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false n-terminal myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false n/a 1537376 unspecified role unspecified role prey prey pyd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false c-terminal egfp tag enhanced green fluorescent protein tag enhanced green fluorescent protein EGFP enhanced green fluorescent protein EGFP egfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false Q9ULZ3:p.Lys21_Lys22delinsGluGlu mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false Q9ULZ3:p.Pro31Arg mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false Q9ULZ3:p.Asp48Arg mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false Q9ULZ3:p.Phe59Glu mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false Q9ULZ3:p.Arg41Glu mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Fig. 6G, Supp. Fig. 6B pycard-casp1 1537375 n/a 1537379 unspecified role unspecified role prey prey n/a 1537376 unspecified role unspecified role bait bait physical association physical association false false false Fig. 7D