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/ Syk is involved in NLRP3 inflammasome-mediated caspase-1 activation through adaptor ASC phosphorylation and enhanced oligomerization. Syk is involved in NLRP3 inflammasome-mediated caspase-1 activation through adaptor ASC phosphorylation and enhanced oligomerization. Journal of leukocyte biology 2015 imex curation Lin YC., Huang DY., Wang JS., Lin YL., Hsieh SL., Huang KC., Lin WW. DSP=82 Fig 3 ASC oliomerisation assay results not added as other mols may be present - no purification (ref 35) wwllaura1119@ntu.edu.tw,bretthuang@ntu.edu.tw 8-Jun-2017: Contacted by IntAct-Help. Only protein-protein interactions mouse-macrophage Mus musculus macrophage macrophage Macrophage anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Lin YC., Huang DY., Wang JS., Lin YL., Hsieh SL., Huang KC., Lin WW. DSP=82 Fig 3 ASC oliomerisation assay results not added as other mols may be present - no purification (ref 35) wwllaura1119@ntu.edu.tw,bretthuang@ntu.edu.tw Accepted 2017-MAY-17 AT 11:47 BST AT 11:47 BST by ORCHARD Journal of leukocyte biology 2015 imex curation I have removed "No UniProt Update" annotation. I am getting a little confused as all the interactions are already shown in Expt-7 but without the No UniProt Update and no Ancillary role for Caspase-1. So can this Expt be deleted?; Added extra interaction- EBI-14036616. Expt-5:-(EBI-14029517) I have removed the "No UniProt Update" annotation at Experiment level, added Myc-ASC as an Ancillary, and NLRP3 as an "agonist" as it is not shown alone. This interaction (EBI-14029517) has been moved to EXpt-4 and this Expt(5) deleted. Syk is involved in NLRP3 inflammasome-mediated caspase-1 activation through adaptor ASC phosphorylation and enhanced oligomerization. Syk is involved in NLRP3 inflammasome-mediated caspase-1 activation through adaptor ASC phosphorylation and enhanced oligomerization. Journal of leukocyte biology 2015 imex curation Lin YC., Huang DY., Wang JS., Lin YL., Hsieh SL., Huang KC., Lin WW. DSP=82 Fig 3 ASC oliomerisation assay results not added as other mols may be present - no purification (ref 35) wwllaura1119@ntu.edu.tw,bretthuang@ntu.edu.tw 8-Jun-2017: Contacted by IntAct-Help. Only protein-protein interactions 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 western blot western blot Immuno blot Immuno blot Lin YC., Huang DY., Wang JS., Lin YL., Hsieh SL., Huang KC., Lin WW. DSP=82 Fig 3 ASC oliomerisation assay results not added as other mols may be present - no purification (ref 35) wwllaura1119@ntu.edu.tw,bretthuang@ntu.edu.tw Accepted 2017-MAY-17 AT 11:47 BST AT 11:47 BST by ORCHARD Journal of leukocyte biology 2015 imex curation I have removed "No UniProt Update" annotation. I am getting a little confused as all the interactions are already shown in Expt-7 but without the No UniProt Update and no Ancillary role for Caspase-1. So can this Expt be deleted?; Added extra interaction- EBI-14036616. Expt-5:-(EBI-14029517) I have removed the "No UniProt Update" annotation at Experiment level, added Myc-ASC as an Ancillary, and NLRP3 as an "agonist" as it is not shown alone. This interaction (EBI-14029517) has been moved to EXpt-4 and this Expt(5) deleted. Syk is involved in NLRP3 inflammasome-mediated caspase-1 activation through adaptor ASC phosphorylation and enhanced oligomerization. Syk is involved in NLRP3 inflammasome-mediated caspase-1 activation through adaptor ASC phosphorylation and enhanced oligomerization. Journal of leukocyte biology 2015 imex curation Lin YC., Huang DY., Wang JS., Lin YL., Hsieh SL., Huang KC., Lin WW. DSP=82 Fig 3 ASC oliomerisation assay results not added as other mols may be present - no purification (ref 35) wwllaura1119@ntu.edu.tw,bretthuang@ntu.edu.tw 8-Jun-2017: Contacted by IntAct-Help. Only protein-protein interactions 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 Lin YC., Huang DY., Wang JS., Lin YL., Hsieh SL., Huang KC., Lin WW. DSP=82 Fig 3 ASC oliomerisation assay results not added as other mols may be present - no purification (ref 35) wwllaura1119@ntu.edu.tw,bretthuang@ntu.edu.tw Accepted 2017-MAY-17 AT 11:47 BST AT 11:47 BST by ORCHARD Journal of leukocyte biology 2015 imex curation I have removed "No UniProt Update" annotation. I am getting a little confused as all the interactions are already shown in Expt-7 but without the No UniProt Update and no Ancillary role for Caspase-1. So can this Expt be deleted?; Added extra interaction- EBI-14036616. Expt-5:-(EBI-14029517) I have removed the "No UniProt Update" annotation at Experiment level, added Myc-ASC as an Ancillary, and NLRP3 as an "agonist" as it is not shown alone. This interaction (EBI-14029517) has been moved to EXpt-4 and this Expt(5) deleted. Syk is involved in NLRP3 inflammasome-mediated caspase-1 activation through adaptor ASC phosphorylation and enhanced oligomerization. Syk is involved in NLRP3 inflammasome-mediated caspase-1 activation through adaptor ASC phosphorylation and enhanced oligomerization. Journal of leukocyte biology 2015 imex curation Lin YC., Huang DY., Wang JS., Lin YL., Hsieh SL., Huang KC., Lin WW. DSP=82 Fig 3 ASC oliomerisation assay results not added as other mols may be present - no purification (ref 35) wwllaura1119@ntu.edu.tw,bretthuang@ntu.edu.tw 8-Jun-2017: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro cleavage cleavage assay cleavage predetermined predetermined participant predetermined Lin YC., Huang DY., Wang JS., Lin YL., Hsieh SL., Huang KC., Lin WW. you (Sandra) said the compound "z-VAD-FMK" could be entered as a (?) 'non-genome entity' or similar, but I cannot find anything remotely similar under "interactor types" - so I have entered it as a "small" molecule. (should be a "medium sized" molecule!) DSP=82 Fig 3 ASC oliomerisation assay results not added as other mols may be present - no purification (ref 35) Caspase activity can be quantified by fluorescent detection of free AFC (also known as 7-amino-4-trifluoromethylcoumarin) CHEBI:136755 wwllaura1119@ntu.edu.tw,bretthuang@ntu.edu.tw Accepted 2017-MAY-17 AT 11:47 BST AT 11:47 BST by ORCHARD Journal of leukocyte biology 2015 imex curation I have removed "No UniProt Update" annotation. I am getting a little confused as all the interactions are already shown in Expt-7 but without the No UniProt Update and no Ancillary role for Caspase-1. So can this Expt be deleted?; Added extra interaction- EBI-14036616. Expt-5:-(EBI-14029517) I have removed the "No UniProt Update" annotation at Experiment level, added Myc-ASC as an Ancillary, and NLRP3 as an "agonist" as it is not shown alone. This interaction (EBI-14029517) has been moved to EXpt-4 and this Expt(5) deleted. Syk is involved in NLRP3 inflammasome-mediated caspase-1 activation through adaptor ASC phosphorylation and enhanced oligomerization. Syk is involved in NLRP3 inflammasome-mediated caspase-1 activation through adaptor ASC phosphorylation and enhanced oligomerization. Journal of leukocyte biology 2015 imex curation Lin YC., Huang DY., Wang JS., Lin YL., Hsieh SL., Huang KC., Lin WW. DSP=82 Fig 3 ASC oliomerisation assay results not added as other mols may be present - no purification (ref 35) wwllaura1119@ntu.edu.tw,bretthuang@ntu.edu.tw 8-Jun-2017: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro protein kinase assay protein kinase assay weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Lin YC., Huang DY., Wang JS., Lin YL., Hsieh SL., Huang KC., Lin WW. DSP=82 Fig 3 ASC oliomerisation assay results not added as other mols may be present - no purification (ref 35) wwllaura1119@ntu.edu.tw,bretthuang@ntu.edu.tw Accepted 2017-MAY-17 AT 11:47 BST AT 11:47 BST by ORCHARD Journal of leukocyte biology 2015 imex curation I have removed "No UniProt Update" annotation. I am getting a little confused as all the interactions are already shown in Expt-7 but without the No UniProt Update and no Ancillary role for Caspase-1. So can this Expt be deleted?; Added extra interaction- EBI-14036616. Expt-5:-(EBI-14029517) I have removed the "No UniProt Update" annotation at Experiment level, added Myc-ASC as an Ancillary, and NLRP3 as an "agonist" as it is not shown alone. This interaction (EBI-14029517) has been moved to EXpt-4 and this Expt(5) deleted. Syk is involved in NLRP3 inflammasome-mediated caspase-1 activation through adaptor ASC phosphorylation and enhanced oligomerization. Syk is involved in NLRP3 inflammasome-mediated caspase-1 activation through adaptor ASC phosphorylation and enhanced oligomerization. Journal of leukocyte biology 2015 imex curation Lin YC., Huang DY., Wang JS., Lin YL., Hsieh SL., Huang KC., Lin WW. DSP=82 Fig 3 ASC oliomerisation assay results not added as other mols may be present - no purification (ref 35) wwllaura1119@ntu.edu.tw,bretthuang@ntu.edu.tw 8-Jun-2017: Contacted by IntAct-Help. Only protein-protein interactions 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 bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Lin YC., Huang DY., Wang JS., Lin YL., Hsieh SL., Huang KC., Lin WW. DSP=82 Fig 3 ASC oliomerisation assay results not added as other mols may be present - no purification (ref 35) wwllaura1119@ntu.edu.tw,bretthuang@ntu.edu.tw Accepted 2017-MAY-17 AT 11:47 BST AT 11:47 BST by ORCHARD Journal of leukocyte biology 2015 imex curation I have removed "No UniProt Update" annotation. I am getting a little confused as all the interactions are already shown in Expt-7 but without the No UniProt Update and no Ancillary role for Caspase-1. So can this Expt be deleted?; Added extra interaction- EBI-14036616. Expt-5:-(EBI-14029517) I have removed the "No UniProt Update" annotation at Experiment level, added Myc-ASC as an Ancillary, and NLRP3 as an "agonist" as it is not shown alone. This interaction (EBI-14029517) has been moved to EXpt-4 and this Expt(5) deleted. asc_mouse Apoptosis-associated speck-like protein containing a CARD Pycard Asc PYD and CARD domain-containing protein protein protein mouse Mus musculus Mouse MGRARDAILDALENLSGDELKKFKMKLLTVQLREGYGRIPRGALLQMDAIDLTDKLVSYYLESYGLELTMTVLRDMGLQELAEQLQTTKEESGAVAAAASVPAQSTARTGHFVDQHRQALIARVTEVDGVLDALHGSVLTEGQYQAVRAETTSQDKMRKLFSFVPSWNLTCKDSLLQALKEIHPYLVMDLEQS DIP protein Q9EPB4 original sequence version: 106 DIP protein Q9EPB4 original sequence version: 104 DIP protein Q9EPB4 original sequence version: 103 2A4EA40194870B31 q96p20-1 NACHT, LRR and PYD domains-containing protein 3 NLRP3 C1orf7 CIAS1 NALP3 PYPAF1 Angiotensin/vasopressin receptor AII/AVP-like Caterpiller protein 1.1 Cold-induced autoinflammatory syndrome 1 protein Cryopyrin PYRIN-containing APAF1-like protein 1 protein protein human Homo sapiens Human MKMASTRCKLARYLEDLEDVDLKKFKMHLEDYPPQKGCIPLPRGQTEKADHVDLATLMIDFNGEEKAWAMAVWIFAAINRRDLYEKAKRDEPKWGSDNARVSNPTVICQEDSIEEEWMGLLEYLSRISICKMKKDYRKKYRKYVRSRFQCIEDRNARLGESVSLNKRYTRLRLIKEHRSQQEREQELLAIGKTKTCESPVSPIKMELLFDPDDEHSEPVHTVVFQGAAGIGKTILARKMMLDWASGTLYQDRFDYLFYIHCREVSLVTQRSLGDLIMSCCPDPNPPIHKIVRKPSRILFLMDGFDELQGAFDEHIGPLCTDWQKAERGDILLSSLIRKKLLPEASLLITTRPVALEKLQHLLDHPRHVEILGFSEAKRKEYFFKYFSDEAQARAAFSLIQENEVLFTMCFIPLVCWIVCTGLKQQMESGKSLAQTSKTTTAVYVFFLSSLLQPRGGSQEHGLCAHLWGLCSLAADGIWNQKILFEESDLRNHGLQKADVSAFLRMNLFQKEVDCEKFYSFIHMTFQEFFAAMYYLLEEEKEGRTNVPGSRLKLPSRDVTVLLENYGKFEKGYLIFVVRFLFGLVNQERTSYLEKKLSCKISQQIRLELLKWIEVKAKAKKLQIQPSQLELFYCLYEMQEEDFVQRAMDYFPKIEINLSTRMDHMVSSFCIENCHRVESLSLGFLHNMPKEEEEEEKEGRHLDMVQCVLPSSSHAACSHGLVNSHLTSSFCRGLFSVLSTSQSLTELDLSDNSLGDPGMRVLCETLQHPGCNIRRLWLGRCGLSHECCFDISLVLSSNQKLVELDLSDNALGDFGIRLLCVGLKHLLCNLKKLWLVSCCLTSACCQDLASVLSTSHSLTRLYVGENALGDSGVAILCEKAKNPQCNLQKLGLVNSGLTSVCCSALSSVLSTNQNLTHLYLRGNTLGDKGIKLLCEGLLHPDCKLQVLELDNCNLTSHCCWDLSTLLTSSQSLRKLSLGNNDLGDLGVMMFCEVLKQQSCLLQNLGLSEMYFNYETKSALETLQEEKPELTVVFEPSW 4C1DFB2B5B283CE8 ac-yvad-afc Ac-YVAD-AFC small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) casp1_mouse Caspase-1 Interleukin-1 beta convertase Interleukin-1 beta-converting enzyme p45 Casp1 Il1bc protein protein mouse Mus musculus Mouse MADKILRAKRKQFINSVSIGTINGLLDELLEKRVLNQEEMDKIKLANITAMDKARDLCDHVSKKGPQASQIFITYICNEDCYLAGILELQSAPSAETFVATEDSKGGHPSSSETKEEQNKEDGTFPGLTGTLKFCPLEKAQKLWKENPSEIYPIMNTTTRTRLALIICNTEFQHLSPRVGAQVDLREMKLLLEDLGYTVKVKENLTALEMVKEVKEFAACPEHKTSDSTFLVFMSHGIQEGICGTTYSNEVSDILKVDTIFQMMNTLKCPSLKDKPKVIIIQACRGEKQGVVLLKDSVRDSEEDFLTDAIFEDDGIKKAHIEKDFIAFCSSTPDNVSWRHPVRGSLFIESLIKHMKEYAWSCDLEDIFRKVRFSFEQPEFRLQMPTADRVTLTKRFYLFPGH DIP protein P29452 original sequence version: 87 DIP protein P29452 original sequence version: 158 DIP protein P29452 original sequence version: 123 DIP protein P29452 original sequence version: 122 3BEBCEFA67C69B03 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 DIP protein Q96P20 original sequence version: 96 DIP protein Q96P20 original sequence version: 160 DIP protein Q96P20-1 original sequence version: 163 DIP protein Q96P20-1 original sequence version: 125 DIP protein Q96P20 original sequence version: 125 4C1DFB2B5B283CE8 ksyk_human Tyrosine-protein kinase SYK SYK Spleen tyrosine kinase p72-Syk protein protein human Homo sapiens Human MASSGMADSANHLPFFFGNITREEAEDYLVQGGMSDGLYLLRQSRNYLGGFALSVAHGRKAHHYTIERELNGTYAIAGGRTHASPADLCHYHSQESDGLVCLLKKPFNRPQGVQPKTGPFEDLKENLIREYVKQTWNLQGQALEQAIISQKPQLEKLIATTAHEKMPWFHGKISREESEQIVLIGSKTNGKFLIRARDNNGSYALCLLHEGKVLHYRIDKDKTGKLSIPEGKKFDTLWQLVEHYSYKADGLLRVLTVPCQKIGTQGNVNFGGRPQLPGSHPATWSAGGIISRIKSYSFPKPGHRKSSPAQGNRQESTVSFNPYEPELAPWAADKGPQREALPMDTEVYESPYADPEEIRPKEVYLDRKLLTLEDKELGSGNFGTVKKGYYQMKKVVKTVAVKILKNEANDPALKDELLAEANVMQQLDNPYIVRMIGICEAESWMLVMEMAELGPLNKYLQQNRHVKDKNIIELVHQVSMGMKYLEESNFVHRDLAARNVLLVTQHYAKISDFGLSKALRADENYYKAQTHGKWPVKWYAPECINYYKFSSKSDVWSFGVLMWEAFSYGQKPYRGMKGSEVTAMLEKGERMGCPAGCPREMYDLMNLCWTYDVENRPGFAAVELRLRNYYYDVVN DIP protein P43405 original sequence version: 146 EAA6BDE65881FC68 nlrp3_mouse NACHT, LRR and PYD domains-containing protein 3 Nlrp3 Cias1 Mmig1 Nalp3 Pypaf1 Cold autoinflammatory syndrome 1 protein homolog Cryopyrin Mast cell maturation-associated-inducible protein 1 PYRIN-containing APAF1-like protein 1 protein protein mouse Mus musculus Mouse MTSVRCKLAQYLEDLEDVDLKKFKMHLEDYPPEKGCIPVPRGQMEKADHLDLATLMIDFNGEEKAWAMAVWIFAAINRRDLWEKAKKDQPEWNDTCTSHSSMVCQEDSLEEEWMGLLGYLSRISICKKKKDYCKMYRRHVRSRFYSIKDRNARLGESVDLNSRYTQLQLVKEHPSKQEREHELLTIGRTKMRDSPMSSLKLELLFEPEDGHSEPVHTVVFQGAAGIGKTILARKIMLDWALGKLFKDKFDYLFFIHCREVSLRTPRSLADLIVSCWPDPNPPVCKILRKPSRILFLMDGFDELQGAFDEHIGEVCTDWQKAVRGDILLSSLIRKKLLPKASLLITTRPVALEKLQHLLDHPRHVEILGFSEAKRKEYFFKYFSNELQAREAFRLIQENEVLFTMCFIPLVCWIVCTGLKQQMETGKSLAQTSKTTTAVYVFFLSSLLQSRGGIEEHLFSDYLQGLCSLAADGIWNQKILFEECDLRKHGLQKTDVSAFLRMNVFQKEVDCERFYSFSHMTFQEFFAAMYYLLEEEAEGETVRKGPGGCSDLLNRDVKVLLENYGKFEKGYLIFVVRFLFGLVNQERTSYLEKKLSCKISQQVRLELLKWIEVKAKAKKLQWQPSQLELFYCLYEMQEEDFVQSAMDHFPKIEINLSTRMDHVVSSFCIKNCHRVKTLSLGFFHNSPKEEEEERRGGRPLDQVQCVFPDTHVACSSRLVNCCLTSSFCRGLFSSLSTNRSLTELDLSDNTLGDPGMRVLCEALQHPGCNIQRLWLGRCGLSHQCCFDISSVLSSSQKLVELDLSDNALGDFGIRLLCVGLKHLLCNLQKLWLVSCCLTSACCQDLALVLSSNHSLTRLYIGENALGDSGVQVLCEKMKDPQCNLQKLGLVNSGLTSICCSALTSVLKTNQNFTHLYLRSNALGDTGLRLLCEGLLHPDCKLQMLELDNCSLTSHSCWNLSTILTHNHSLRKLNLGNNDLGDLCVVTLCEVLKQQGCLLQSLQLGEMYLNRETKRALEALQEEKPELTIVFEISW DIP protein Q8R4B8 original sequence version: 102 DIP protein Q8R4B8 original sequence version: 134 DIP protein Q8R4B8 original sequence version: 99 5924690966B12117 z-vad-fmk small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) 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 DIP protein Q9ULZ3 original sequence version: 116 455987286586F46A p29466-pro_0000004522 Caspase-1 subunit p20 CASP1 IL1BC IL1BCE Interleukin-1 beta convertase Interleukin-1 beta-converting enzyme p45 protein protein human Homo sapiens Human NPAMPTSSGSEGNVKLCSLEEAQRIWKQKSAEIYPIMDKSSRTRLALIICNEEFDSIPRRTGAEVDITGMTMLLQNLGYSVDVKKNLTASDMTTELEAFAHRPEHKTSDSTFLVFMSHGIREGICGKKHSEQVPDILQLNAIFNMLNTKNCPSLKDKPKVIIIQACRGDSPGVVWFKD 120 297 6E7413E046F7629E p29466-pro_0000004524 Caspase-1 subunit p10 CASP1 IL1BC IL1BCE Interleukin-1 beta convertase Interleukin-1 beta-converting enzyme p45 protein protein human Homo sapiens Human AIKKAHIEKDFIAFCSSTPDNVSWRHPTMGSVFIGRLIEHMQEYACSCDVEEIFRKVRFSFEQPDGRAQMPTTERVTLTRCFYLFPGH 317 404 1130E0FB030FF748 casp1_human Caspase-1 Interleukin-1 beta convertase Interleukin-1 beta-converting enzyme p45 CASP1 IL1BC IL1BCE protein protein human Homo sapiens Human MADKVLKEKRKLFIRSMGEGTINGLLDELLQTRVLNKEEMEKVKRENATVMDKTRALIDSVIPKGAQACQICITYICEEDSYLAGTLGLSADQTSGNYLNMQDSQGVLSSFPAPQAVQDNPAMPTSSGSEGNVKLCSLEEAQRIWKQKSAEIYPIMDKSSRTRLALIICNEEFDSIPRRTGAEVDITGMTMLLQNLGYSVDVKKNLTASDMTTELEAFAHRPEHKTSDSTFLVFMSHGIREGICGKKHSEQVPDILQLNAIFNMLNTKNCPSLKDKPKVIIIQACRGDSPGVVWFKDSVGVSGNLSLPTTEEFEDDAIKKAHIEKDFIAFCSSTPDNVSWRHPTMGSVFIGRLIEHMQEYACSCDVEEIFRKVRFSFEQPDGRAQMPTTERVTLTRCFYLFPGH DIP protein P29466 original sequence version: 155 DIP protein P29466-1 original sequence version: 169 DIP protein P29466 original sequence version: 169 DIP protein P29466 original sequence version: 163 DIP protein P29466 original sequence version: 101 DIP protein P29466 original sequence version: 160 DIP protein P29466 original sequence version: 184 ABF33CF33CC71584 ksyk_mouse Tyrosine-protein kinase SYK Syk Spleen tyrosine kinase Sykb ptk72 protein protein mouse Mus musculus Mouse MAGSAVDSANHLTYFFGNITREEAEDYLVQGGMTDGLYLLRQSRNYLGGFALSVAHNRKAHHYTIERELNGTYAISGGRAHASPADLCHYHSQEPDGLICLLKKPFNRPPGVQPKTGPFEDLKENLIREYVKQTWNLQGQALEQAIISQKPQLEKLIATTAHEKMPWFHGNISRDESEQTVLIGSKTNGKFLIRARDNSGSYALCLLHEGKVLHYRIDRDKTGKLSIPEGKKFDTLWQLVEHYSYKPDGLLRVLTVPCQKIGAQMGHPGSPNAHPVTWSPGGIISRIKSYSFPKPGHKKPAPPQGSRPESTVSFNPYEPTGGPWGPDRGLQREALPMDTEVYESPYADPEEIRPKEVYLDRSLLTLEDNELGSGNFGTVKKGYYQMKKVVKTVAVKILKNEANDPALKDELLAEANVMQQLDNPYIVRMIGICEAESWMLVMEMAELGPLNKYLQQNRHIKDKNIIELVHQVSMGMKYLEESNFVHRDLAARNVLLVTQHYAKISDFGLSKALRADENYYKAQTHGKWPVKWYAPECINYYKFSSKSDVWSFGVLMWEAFSYGQKPYRGMKGSEVTAMLEKGERMGCPAGCPREMYDLMNLCWTYDVENRPGFTAVELRLRNYYYDVVN DIP protein P48025 original sequence version: 108 DIP protein P48025 original sequence version: 125 2F98D21601F8224E ac_yvad_afc-pro_0000004522_pro_0000004524-1 4 9 enzyme target enzyme target substrate substrate neutral component neutral component 17 enzyme enzyme neutral component neutral component whole_molecule sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 16 enzyme enzyme neutral component neutral component whole_molecule sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 23 25 cleavage cleavage reaction cleavage 1F 5arhsss-1 1 7 unspecified role unspecified role prey prey 10 unspecified role unspecified role bait bait physical association physical association 4B, Supp 2B The interaction between ASC and caspase-1 under ATP and Alum treatment was reduced markedly in the Syk−/− macrophages. ATP Alum adjuvant (aluminum hydroxide and magnesium hydroxide) casp1-pycard-2 1 7 unspecified role unspecified role bait bait 19 unspecified role unspecified role prey prey 13 unspecified role unspecified role prey prey association association 4A ATP With the use of ASC antibody for immunoprecipitation, we also detected the association of ASC to Syk and NLRP3 following ATP treatment (Fig. 4A, right). ac_yvad_afc-casp1-1 4 14 inhibitor inhibitor neutral component neutral component 9 enzyme target enzyme target substrate substrate neutral component neutral component 17 enzyme enzyme neutral component neutral component whole_molecule sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 16 enzyme enzyme neutral component neutral component whole_molecule sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 39 37 cleavage cleavage reaction cleavage 1Fi syk-nlrp3-1 3 11 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 12 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined dSH2_construct sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain kinase_domain required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain physical association physical association 4C casp1-syk-1 2 15 unspecified role unspecified role ancillary ancillary region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q9ULZ3:p.Tyr146Phe mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain Y F Wed May 02 16:43:25 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr146phe' Tue Jun 30 19:17:32 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr146Phe' Q9ULZ3:p.Tyr187Phe mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain Y F Wed May 02 19:40:36 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr187phe' Tue Jun 30 15:56:41 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr187Phe' 12 unspecified role unspecified role prey prey 18 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association 4 E/F, 5C Cells expressed Myc-tagged ASC and NLRP3 OR Myc-ASC alone. No direct interaction was observed between Syk and procaspase-1. NLRP3 syk-nlrp3-2 2 12 unspecified role unspecified role bait bait dSH2_construct sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain kinase_domain required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 15 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association 4D 233467474-1 5 12 putative self putative self neutral component neutral component region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined phosphorylation phosphorylation reaction phosphorylation true 5 A syk-8 5 12 enzyme enzyme neutral component neutral component region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 15 enzyme target enzyme target substrate substrate neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain Q9ULZ3:p.Ser164Asn mutation mutation certain certain sequence position certain certain certain sequence position certain S N Wed May 02 19:11:24 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser164asn' Tue Jun 30 19:06:01 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser164Asn' in vitro In vitro phosphorylation phosphorylation reaction phosphorylation 5 A P43405 (GO:0004672) up-regulates activity of Q9ULZ3 (GO:0051259) As a result, we found that the ASC protein of Y146 or Y187 mutation remained susceptible to phosphorylation by Syk, whereas the double mutations of ASC Y146/187F robustly reduced the extent of Syk-dependent phosphorylation (Fig. 5B). nlrp3-syk-1 5 12 enzyme enzyme neutral component neutral component region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 enzyme target enzyme target substrate substrate neutral component neutral component n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range in vitro In vitro phosphorylation phosphorylation reaction phosphorylation 5 A P43405 (GO:0004672) up-regulates activity of Q96P20-1 (GO:1900225) casp1-syk-2 6 12 unspecified role unspecified role prey prey 18 anti tag western anti tag western blot anti tag western western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 15 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association 5D pycard-casp1-1 6 18 anti tag western anti tag western blot anti tag western western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 15 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q9ULZ3:p.Tyr146Phe mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain Y F Wed May 02 20:16:40 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr146phe' Tue Jun 30 19:55:50 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr146Phe' Q9ULZ3:p.Tyr187Phe mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain Y F Wed May 02 16:34:06 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr187phe' Tue Jun 30 16:17:53 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr187Phe' Q9ULZ3:p.[Tyr146Phe;Tyr187Phe] mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain Y F certain certain sequence position certain certain certain sequence position certain Y F Wed May 02 14:25:27 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr146phe,tyr187phe' Tue Jun 30 19:45:03 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Tyr146Phe;Tyr187Phe]' physical association physical association 5Di pycard-casp1-2 6 18 unspecified role unspecified role ancillary ancillary 12 anti tag western anti tag western blot anti tag western western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey 15 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q9ULZ3:p.Tyr146Phe mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain Y F Wed May 02 14:41:41 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr146phe' Tue Jun 30 17:06:26 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr146Phe' Q9ULZ3:p.Tyr187Phe mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain Y F Wed May 02 19:08:24 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr187phe' Tue Jun 30 18:05:53 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Tyr187Phe' Q9ULZ3:p.[Tyr146Phe;Tyr187Phe] mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain Y F certain certain sequence position certain certain certain sequence position certain Y F Wed May 02 16:42:54 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr146phe,tyr187phe' Tue Jun 30 15:56:20 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Tyr146Phe;Tyr187Phe]' association association 5Dii Only WT but not mutant (Y146F and/or Y187F) ASC can trigger the binding between Syk and procaspase-1 Conversely, with the use of ASC antibody for immunoprecipitation, we also achieved similar results, i.e., only WT but not mutant ASC can trigger the binding between Syk and procaspase-1 (Fig. 5D) casp1-pycard-3 1 19 unspecified role unspecified role bait bait 10 unspecified role unspecified role bait bait physical association physical association Supp 2A ATP Alum adjuvant (aluminum hydroxide and magnesium hydroxide) In addition, we observed the association of Syk with procaspase-1 upon ATP or Alum treatment (Supplemental Fig. 2A). pycard-nlrp3-1 1 7 unspecified role unspecified role prey prey 19 unspecified role unspecified role bait bait 13 unspecified role unspecified role prey prey association association 4A-2 ATP We performed immunoprecipitation experiments by use of Syk antibody and observed the association of Syk with ASC and NLRP3 after ATP stimulation for 20 min (Fig. 4A, middle).