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/ bonham-2014-3 A promiscuous lipid-binding protein diversifies the subcellular sites of toll-like receptor signal transduction. in vitro In vitro far western blotting far western blotting Affinity blotting Affinity blotting western blot western blot Immuno blot Immuno blot Bonham K.S., Orzalli M.H., Hayashi K., Wolf A.I., Glanemann C., Weninger W., Iwasaki A., Knipe D.M., Kagan J.C. Details of GST-TIRAP purification adn purity taken from the referenced article, PMID: 16751103 jonathan.kagan@childrens.harvard.edu Cell (0092-8674) 2014 Only protein-protein interactions imex curation bonham-2014-1 A promiscuous lipid-binding protein diversifies the subcellular sites of toll-like receptor signal transduction. mouse-bmdm Mus musculus bmdm bone marrow-derived macrophage anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Bonham K.S., Orzalli M.H., Hayashi K., Wolf A.I., Glanemann C., Weninger W., Iwasaki A., Knipe D.M., Kagan J.C. jonathan.kagan@childrens.harvard.edu Cell (0092-8674) 2014 Only protein-protein interactions imex curation bonham-2014-2 A promiscuous lipid-binding protein diversifies the subcellular sites of toll-like receptor signal transduction. mouse-bmdm Mus musculus bmdm bone marrow-derived macrophage pull down pull down western blot western blot Immuno blot Immuno blot Bonham K.S., Orzalli M.H., Hayashi K., Wolf A.I., Glanemann C., Weninger W., Iwasaki A., Knipe D.M., Kagan J.C. jonathan.kagan@childrens.harvard.edu Cell (0092-8674) 2014 Only protein-protein interactions imex curation tirap_human Toll/interleukin-1 receptor domain-containing adapter protein MyD88 adapter-like protein Adaptor protein Wyatt TIRAP MAL protein protein human Homo sapiens Human MASSTSLPAPGSRPKKPLGKMADWFRQTLLKKPKKRPNSPESTSSDASQPTSQDSPLPPSLSSVTSPSLPPTHASDSGSSRWSKDYDVCVCHSEEDLVAAQDLVSYLEGSTASLRCFLQLRDATPGGAIVSELCQALSSSHCRVLLITPGFLQDPWCKYQMLQALTEAPGAEGCTIPLLSGLSRAAYPPELRFMYYVDGRGPDGGFRQVKEAVMRYLQTLS pi3p 1-phosphatidyl-1D-myo-inositol 3-phosphates small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) pi4p 1-phosphatidyl-1D-myo-inositol 4-phosphate small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) pi5p 1-phosphatidyl-1D-myo-inositol 5-phosphate small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) 1-phosphatidyl-1d-myo-inositol 3,4-bisphosphate 1-phosphatidyl-1D-myo-inositol 3,4-bisphosphate small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) 1-phosphatidyl-1d-myo-inositol 3,5-bisphosphate 1-phosphatidyl-1D-myo-inositol 3,5-bisphosphates small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) 1-phosphatidyl-1d-myo-inositol 3,4,5-trisphosphate 1-phosphatidyl-1D-myo-inositol 3,4,5-trisphosphates small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) 1-phosphatidyl-1d-myo-inositol 4,5-bisphosphate 1-phosphatidyl-1D-myo-inositol 4,5-bisphosphates small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) phosphatidyl-l-serines phosphatidyl-L-serines small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) irak4_mouse Interleukin-1 receptor-associated kinase 4 Irak4 protein protein mouse Mus musculus Mouse MNKPLTPSTYIRNLNVGILRKLSDFIDPQEGWKKLAVAIKKPSGDDRYNQFHIRRFEALLQTGKSPTCELLFDWGTTNCTVGDLVDLLVQIELFAPATLLLPDAVPQTVKSLPPREAATVAQTHGPCQEKDRTSVMPMPKLEHSCEPPDSSSPDNRSVESSDTRFHSFSFHELKSITNNFDEQPASAGGNRMGEGGFGVVYKGCVNNTIVAVKKLGAMVEISTEELKQQFDQEIKVMATCQHENLVELLGFSSDSDNLCLVYAYMPNGSLLDRLSCLDGTPPLSWHTRCKVAQGTANGIRFLHENHHIHRDIKSANILLDKDFTAKISDFGLARASARLAQTVMTSRIVGTTAYMAPEALRGEITPKSDIYSFGVVLLELITGLAAVDENREPQLLLDIKEEIEDEEKTIEDYTDEKMSDADPASVEAMYSAASQCLHEKKNRRPDIAKVQQLLQEMSA myd88_mouse Myeloid differentiation primary response protein MyD88 Myd88 protein protein mouse Mus musculus Mouse MSAGDPRVGSGSLDSFMFSIPLVALNVGVRRRLSLFLNPRTPVAADWTLLAEEMGFEYLEIRELETRPDPTRSLLDAWQGRSGASVGRLLELLALLDREDILKELKSRIEEDCQKYLGKQQNQESEKPLQVARVESSVPQTKELGGITTLDDPLGQTPELFDAFICYCPNDIEFVQEMIRQLEQTDYRLKLCVSDRDVLPGTCVWSIASELIEKRCRRMVVVVSDDYLQSKECDFQTKFALSLSPGVQQKRLIPIKYKAMKKDFPSILRFITICDYTNPCTKSWFWTRLAKALSLP irak2_mouse Interleukin-1 receptor-associated kinase-like 2 Irak2 protein protein mouse Mus musculus Mouse MACYIYQLPSWVLDDLCRNIDTLSEWDWMQFASYVITDLTQLRKIKSMERVQGVSITRELLWWWSMRQATVQQLVDLLCHLELYRAAQIVLSWKPVPESTSPLPAFPEAVKPGAVATSRRNLKDEQEKVRPVKPRSLLDTGPIMAGAQRQRPCEMDAPCSLKTDAPDSPQSKYCSTSTSAPKQERLLGLPGDRLFWSEADVVQATEDFDQSHRISEGTFADIYQGQRNGVAFAFKKLREVAGSSPGSMDRFLQAEMQLCLRCCHANVLPLLGFCTGRQFHSLIYPYMANGSLHDRLWAQGNSDMLPWPQRASICSGLLLAVEHLHSLDIIHSNVKSANVLLDQHLNPKLAHPVAHPHPDNKKTKYTVMRTHLFQASAAYLPEHFIRVGQLTKQVDIFSCGIVLAEVLTGIPAMDKDRSPVYLKDLLLSEIPNSTSSVCSRKTSMGKAVVKEICQRHVEKRAGLLPEACEEAWATAVSVCLRRRNASVEEARVSLAGVEEQLRGQLSLPWSRVSEATGSSSNTPEETDDVDNSSLSVPSLVMMASCPGAASSPLFTGHGAAQPSTSGRQEADSSSEACTGPQTPQNATETSWKIEINEAKRRLMENIVLYKEERLDSSELFGP pi3p-tirap 1535254 n/a 1535257 unspecified role unspecified role prey prey 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 false P58753:p.[Lys15_Lys16delinsAlaAla;Lys31_Lys32delinsAlaAla] mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1535258 unspecified role unspecified role bait bait direct interaction direct interaction false false false pi4p-tirap 1535254 n/a 1535257 unspecified role unspecified role prey prey P58753:p.[Lys15_Lys16delinsAlaAla;Lys31_Lys32delinsAlaAla] mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false 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 false ecoli Escherichia coli (strain K12) n/a 1535259 unspecified role unspecified role bait bait direct interaction direct interaction false false false Fig. 4A pi5p-tirap 1535254 n/a 1535257 unspecified role unspecified role prey prey P58753:p.[Lys15_Lys16delinsAlaAla;Lys31_Lys32delinsAlaAla] mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false 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 false ecoli Escherichia coli (strain K12) n/a 1535260 unspecified role unspecified role bait bait direct interaction direct interaction false false false Fig. 4A 1_phosphatidyl_1d_myo_inositol_3_4_bisphosphate-tirap 1535254 n/a 1535257 unspecified role unspecified role prey prey P58753:p.[Lys15_Lys16delinsAlaAla;Lys31_Lys32delinsAlaAla] mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false 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 false ecoli Escherichia coli (strain K12) n/a 1535261 unspecified role unspecified role bait bait direct interaction direct interaction false false false Fig. 4A 1_phosphatidyl_1d_myo_inositol_3_5_bisphosphate-tirap 1535254 n/a 1535257 unspecified role unspecified role prey prey P58753:p.[Lys15_Lys16delinsAlaAla;Lys31_Lys32delinsAlaAla] mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false 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 false ecoli Escherichia coli (strain K12) n/a 1535262 unspecified role unspecified role bait bait direct interaction direct interaction false false false Fig. 4A 1_phosphatidyl_1d_myo_inositol_3_4_5_trisphosphate-tirap 1535254 n/a 1535257 unspecified role unspecified role prey prey 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 false P58753:p.[Lys15_Lys16delinsAlaAla;Lys31_Lys32delinsAlaAla] mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1535263 unspecified role unspecified role bait bait direct interaction direct interaction false false false Fig. 4A 1_phosphatidyl_1d_myo_inositol_4_5_bisphosphate-tirap 1535254 n/a 1535257 unspecified role unspecified role prey prey P58753:p.[Lys15_Lys16delinsAlaAla;Lys31_Lys32delinsAlaAla] mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false 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 false ecoli Escherichia coli (strain K12) n/a 1535264 unspecified role unspecified role bait bait direct interaction direct interaction false false false Fig. 4A phosphatidyl_l_serines-tirap 1535254 n/a 1535257 unspecified role unspecified role prey prey P58753:p.[Lys15_Lys16delinsAlaAla;Lys31_Lys32delinsAlaAla] mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false 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 false ecoli Escherichia coli (strain K12) n/a 1535265 unspecified role unspecified role bait bait direct interaction direct interaction false false false Fig. 4A myd88-irak2 1535255 n/a 1535266 unspecified role unspecified role prey prey n/a 1535267 unspecified role unspecified role bait bait n/a 1535268 unspecified role unspecified role prey prey association association false false false Fig. 3A, 3B Lipopolysaccharide CpG DNA irak2-irak4 1535255 n/a 1535266 unspecified role unspecified role prey prey n/a 1535268 unspecified role unspecified role bait bait n/a 1535267 unspecified role unspecified role prey prey association association false false false Supp. Fig. 2A Lipopolysaccharide CpG DNA myd88-irak2-3 1535255 n/a 1535266 unspecified role unspecified role prey prey n/a 1535267 unspecified role unspecified role bait bait physical association physical association false false false Fig.3E, 5C TIRAP -/- cells reconstituted withwild-type TIRAP formed myddosomes in response to both lipopolysaccharide (LPS) and CpG DNA. By contrast, PLC-TIRAP was able to rescue myddosome formation solely in response to LPS, whereas PX-TIRAP and SLP2a-TIRAP were able to rescue myddosome formation in response to CpG DNA, but not LPS Lipopolysaccharide CpG DNA myd88-irak2-2 1535255 n/a 1535266 unspecified role unspecified role prey prey n/a 1535268 unspecified role unspecified role prey prey n/a 1535257 anti tag western anti tag western blot anti tag western unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1535267 unspecified role unspecified role bait bait association association false false false Supp. Fig.2B myd88-irak2-4 1535255 n/a 1535266 unspecified role unspecified role prey prey n/a 1535268 unspecified role unspecified role prey prey n/a 1535257 anti tag western anti tag western blot anti tag western unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1535267 unspecified role unspecified role bait bait association association false false false Supp. Fig. 2B tirap-irak4 1535256 n/a 1535266 unspecified role unspecified role prey prey n/a 1535267 unspecified role unspecified role prey prey n/a 1535257 unspecified role unspecified role bait bait region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false association association false false false Figs. 3C, 3D LPS, CpG DNA