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/ sakaguchi-2010-5 human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Sakaguchi M., Murata H., Yamamoto K., Ono T., Sakaguchi Y., Motoyama A., Hibino T., Kataoka K., Huh NH. Where both prey proteins were myc-tagged, they were further identified by molecular weight. Tirap and MyD88 are assumed to be human as they are same construct as those used in Fig. 2D where Figure legend 2 says: (D and E) Abortive signal transduction from activated endogenous RAGE in MyD88-/- or TIRAP-down regulated mouse fibroblasts was rescued by forced expression of the corresponding human genes. Also according to M&M they have N-terminal Myc and C-terminal HA. Authors provided origin species of recombinant PKCzeta as human namu@md.okayama-u.ac.jp PloS one 2010 imex curation Only protein-protein interactions imex curation sakaguchi-2010-1 human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Sakaguchi M., Murata H., Yamamoto K., Ono T., Sakaguchi Y., Motoyama A., Hibino T., Kataoka K., Huh NH. namu@md.okayama-u.ac.jp PloS one 2010 imex curation Only protein-protein interactions imex curation sakaguchi-2010-3 mouse-fibroblast Mus musculus fibroblast Fibroblast Resident cell of connective tissue, mesodermally derived anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Sakaguchi M., Murata H., Yamamoto K., Ono T., Sakaguchi Y., Motoyama A., Hibino T., Kataoka K., Huh NH. endogenous MyD88 is knocked down Figure legend 2 says: (D and E) Abortive signal transduction from activated endogenous RAGE in MyD88-/- or TIRAP-down regulated mouse fibroblasts was rescued by forced expression of the corresponding human genes. So RAGE is endogenous and TIRAP and MyD88 are human when over expressed. Also according to M&M they have N-terminal Myc and C-terminal HA. namu@md.okayama-u.ac.jp PloS one 2010 imex curation Only protein-protein interactions imex curation sakaguchi-2010-4 mouse-nih_3t3 Mus musculus Swiss NIH 3t3 embryo fibroblast cells nih_3t3 Mouse Swiss NIH embryo contact inhibited fibroblast cells anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Sakaguchi M., Murata H., Yamamoto K., Ono T., Sakaguchi Y., Motoyama A., Hibino T., Kataoka K., Huh NH. endogenous TIRAP is silenced Figure legend 2 says: (D and E) Abortive signal transduction from activated endogenous RAGE in MyD88-/- or TIRAP-down regulated mouse fibroblasts was rescued by forced expression of the corresponding human genes. So RAGE is endogenous and TIRAP and MyD88 are human when over expressed. Also according to M&M they have N-terminal Myc and C-terminal HA. namu@md.okayama-u.ac.jp PloS one 2010 imex curation Only protein-protein interactions imex curation sakaguchi-2010-6 human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Sakaguchi M., Murata H., Yamamoto K., Ono T., Sakaguchi Y., Motoyama A., Hibino T., Kataoka K., Huh NH. 293-hMD2-CD14 cells used (stably expressing human CD14 and human MD2) TLR-4 has been inferred to be of human origing, according to following sentence:  "and in the cytoplasmic domain of TLR4 (QDED starting at 683)", which corresponds to human protein. namu@md.okayama-u.ac.jp PloS one 2010 imex curation Only protein-protein interactions imex curation sakaguchi-2010-2 human-uvec Human Umbilical Vein Endothelial Cells (HUVEC) huvec umbilical_vein_endothelium Umbilical vein endothelial cell [Umbilical vein endothelium] anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Sakaguchi M., Murata H., Yamamoto K., Ono T., Sakaguchi Y., Motoyama A., Hibino T., Kataoka K., Huh NH. namu@md.okayama-u.ac.jp PloS one 2010 imex curation 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 rage_human Advanced glycosylation end product-specific receptor AGER RAGE Receptor for advanced glycosylation end products protein protein human Homo sapiens Human MAAGTAVGAWVLVLSLWGAVVGAQNITARIGEPLVLKCKGAPKKPPQRLEWKLNTGRTEAWKVLSPQGGGPWDSVARVLPNGSLFLPAVGIQDEGIFRCQAMNRNGKETKSNYRVRVYQIPGKPEIVDSASELTAGVPNKVGTCVSEGSYPAGTLSWHLDGKPLVPNEKGVSVKEQTRRHPETGLFTLQSELMVTPARGGDPRPTFSCSFSPGLPRHRALRTAPIQPRVWEPVPLEEVQLVVEPEGGAVAPGGTVTLTCEVPAQPSPQIHWMKDGVPLPLPPSPVLILPEIGPQDQGTYSCVATHSSHGPQESRAVSISIIEPGEEGPTAGSVGGSGLGTLALALGILGGLGTAALLIGVILWQRRQRRGEERKAPENQEEEEERAELNQSEEPEAGESSTGGP myd88_human Myeloid differentiation primary response protein MyD88 MYD88 protein protein human Homo sapiens Human MAAGGPGAGSAAPVSSTSSLPLAALNMRVRRRLSLFLNVRTQVAADWTALAEEMDFEYLEIRQLETQADPTGRLLDAWQGRPGASVGRLLELLTKLGRDDVLLELGPSIEEDCQKYILKQQQEEAEKPLQVAAVDSSVPRTAELAGITTLDDPLGHMPERFDAFICYCPSDIQFVQEMIRQLEQTNYRLKLCVSDRDVLPGTCVWSIASELIEKRCRRMVVVVSDDYLQSKECDFQTKFALSLSPGAHQKRLIPIKYKAMKKEFPSILRFITVCDYTNPCTKSWFWTRLAKALSLP kpcz_human Protein kinase C zeta type PRKCZ PKC2 nPKC-zeta protein protein human Homo sapiens Human MPSRTGPKMEGSGGRVRLKAHYGGDIFITSVDAATTFEELCEEVRDMCRLHQQHPLTLKWVDSEGDPCTVSSQMELEEAFRLARQCRDEGLIIHVFPSTPEQPGLPCPGEDKSIYRRGARRWRKLYRANGHLFQAKRFNRRAYCGQCSERIWGLARQGYRCINCKLLVHKRCHGLVPLTCRKHMDSVMPSQEPPVDDKNEDADLPSEETDGIAYISSSRKHDSIKDDSEDLKPVIDGMDGIKISQGLGLQDFDLIRVIGRGSYAKVLLVRLKKNDQIYAMKVVKKELVHDDEDIDWVQTEKHVFEQASSNPFLVGLHSCFQTTSRLFLVIEYVNGGDLMFHMQRQRKLPEEHARFYAAEICIALNFLHERGIIYRDLKLDNVLLDADGHIKLTDYGMCKEGLGPGDTTSTFCGTPNYIAPEILRGEEYGFSVDWWALGVLMFEMMAGRSPFDIITDNPDMNTEDYLFQVILEKPIRIPRFLSVKASHVLKGFLNKDPKERLGCRPQTGFSDIKSHAFFRSIDWDLLEKKQALPPFQPQITDDYGLDNFDTQFTSEPVQLTPDDEDAIKRIDQSEFEGFEYINPLLLSTEESV irak4_human Interleukin-1 receptor-associated kinase 4 Renal carcinoma antigen NY-REN-64 IRAK4 protein protein human Homo sapiens Human MNKPITPSTYVRCLNVGLIRKLSDFIDPQEGWKKLAVAIKKPSGDDRYNQFHIRRFEALLQTGKSPTSELLFDWGTTNCTVGDLVDLLIQNEFFAPASLLLPDAVPKTANTLPSKEAITVQQKQMPFCDKDRTLMTPVQNLEQSYMPPDSSSPENKSLEVSDTRFHSFSFYELKNVTNNFDERPISVGGNKMGEGGFGVVYKGYVNNTTVAVKKLAAMVDITTEELKQQFDQEIKVMAKCQHENLVELLGFSSDGDDLCLVYVYMPNGSLLDRLSCLDGTPPLSWHMRCKIAQGAANGINFLHENHHIHRDIKSANILLDEAFTAKISDFGLARASEKFAQTVMTSRIVGTTAYMAPEALRGEITPKSDIYSFGVVLLEIITGLPAVDEHREPQLLLDIKEEIEDEEKTIEDYIDKKMNDADSTSVEAMYSVASQCLHEKKNKRPDIKKVQQLLQEMTAS irak4_mouse Interleukin-1 receptor-associated kinase 4 Irak4 protein protein mouse Mus musculus Mouse MNKPLTPSTYIRNLNVGILRKLSDFIDPQEGWKKLAVAIKKPSGDDRYNQFHIRRFEALLQTGKSPTCELLFDWGTTNCTVGDLVDLLVQIELFAPATLLLPDAVPQTVKSLPPREAATVAQTHGPCQEKDRTSVMPMPKLEHSCEPPDSSSPDNRSVESSDTRFHSFSFHELKSITNNFDEQPASAGGNRMGEGGFGVVYKGCVNNTIVAVKKLGAMVEISTEELKQQFDQEIKVMATCQHENLVELLGFSSDSDNLCLVYAYMPNGSLLDRLSCLDGTPPLSWHTRCKVAQGTANGIRFLHENHHIHRDIKSANILLDKDFTAKISDFGLARASARLAQTVMTSRIVGTTAYMAPEALRGEITPKSDIYSFGVVLLELITGLAAVDENREPQLLLDIKEEIEDEEKTIEDYTDEKMSDADPASVEAMYSAASQCLHEKKNRRPDIAKVQQLLQEMSA kpcz_mouse Protein kinase C zeta type nPKC-zeta Prkcz Pkcz protein protein mouse Mus musculus Mouse MPSRTDPKMDRSGGRVRLKAHYGGDILITSVDAMTTFKDLCEEVRDMCGLHQQHPLTLKWVDSEGDPCTVSSQMELEEAFRLVCQGRDEVLIIHVFPSIPEQPGMPCPGEDKSIYRRGARRWRKLYRANGHLFQAKRFNRGAYCGQCSERIWGLSRQGYRCINCKLLVHKRCHVLVPLTCRRHMDSVMPSQEPPVDDKNDGVDLPSEETDGIAYISSSRKHDNIKDDSEDLKPVIDGVDGIKISQGLGLQDFDLIRVIGRGSYAKVLLVRLKKNDQIYAMKVVKKELVHDDEDIDWVQTEKHVFEQASSNPFLVGLHSCFQTTSRLFLVIEYVNGGDLMFHMQRQRKLPEEHARFYAAEICIALNFLHERGIIYRDLKLDNVLLDADGHIKLTDYGMCKEGLGPGDTTSTFCGTPNYIAPEILRGEEYGFSVDWWALGVLMFEMMAGRSPFDIITDNPDMNTEDYLFQVILEKPIRIPRFLSVKASHVLKGFLNKDPKERLGCRPQTGFSDIKSHAFFRSIDWDLLEKKQTLPPFQPQITDDYGLDNFDTQFTSEPVQLTPDDEDVIKRIDQSEFEGFEYINPLLLSAEESV rage_mouse Advanced glycosylation end product-specific receptor Ager Rage Receptor for advanced glycosylation end products protein protein mouse Mus musculus Mouse MPAGTAARAWVLVLALWGAVAGGQNITARIGEPLVLSCKGAPKKPPQQLEWKLNTGRTEAWKVLSPQGGPWDSVARILPNGSLLLPATGIVDEGTFRCRATNRRGKEVKSNYRVRVYQIPGKPEIVDPASELTASVPNKVGTCVSEGSYPAGTLSWHLDGKLLIPDGKETLVKEETRRHPETGLFTLRSELTVIPTQGGTHPTFSCSFSLGLPRRRPLNTAPIQLRVREPGPPEGIQLLVEPEGGIVAPGGTVTLTCAISAQPPPQVHWIKDGAPLPLAPSPVLLLPEVGHEDEGTYSCVATHPSHGPQESPPVSIRVTETGDEGPAEGSVGESGLGTLALALGILGGLGVVALLVGAILWRKRQPRREERKAPESQEDEEERAELNQSEEAEMPENGAGGP tirap_mouse Toll/interleukin-1 receptor domain-containing adapter protein Tirap Mal Adaptor protein Wyatt MyD88 adapter-like protein protein protein mouse Mus musculus Mouse MASSSSVPASSTPSKKPRDKIADWFRQALLKKPKKMPISQESHLYDGSQTATQDGLSPSSCSSPPSHSSPESRSSPSSCSSGMSPTSPPTHVDSSSSSSGRWSKDYDVCVCHSEEDLEAAQELVSYLEGSQASLRCFLQLRDAAPGGAIVSELCQALSRSHCRALLITPGFLRDPWCKYQMLQALTEAPASEGCTIPLLSGLSRAAYPPELRFMYYVDGRGKDGGFYQVKEAVIHYLETLS myd88_mouse Myeloid differentiation primary response protein MyD88 Myd88 protein protein mouse Mus musculus Mouse MSAGDPRVGSGSLDSFMFSIPLVALNVGVRRRLSLFLNPRTPVAADWTLLAEEMGFEYLEIRELETRPDPTRSLLDAWQGRSGASVGRLLELLALLDREDILKELKSRIEEDCQKYLGKQQNQESEKPLQVARVESSVPQTKELGGITTLDDPLGQTPELFDAFICYCPNDIEFVQEMIRQLEQTDYRLKLCVSDRDVLPGTCVWSIASELIEKRCRRMVVVVSDDYLQSKECDFQTKFALSLSPGVQQKRLIPIKYKAMKKDFPSILRFITICDYTNPCTKSWFWTRLAKALSLP tcam2_human TIR domain-containing adapter molecule 2 TRIF-related adapter molecule Toll/interleukin-1 receptor domain-containing protein Toll-like receptor adaptor protein 3 Putative NF-kappa-B-activating protein 502 TICAM2 TIRP TIRAP3 TRAM protein protein human Homo sapiens Human MGIGKSKINSCPLSLSWGKRHSVDTSPGYHESDSKKSEDLSLCNVAEHSNTTEGPTGKQEGAQSVEEMFEEEAEEEVFLKFVILHAEDDTDEALRVQNLLQDDFGIKPGIIFAEMPCGRQHLQNLDDAVNGSAWTILLLTENFLRDTWCNFQFYTSLMNSVNRQHKYNSVIPMRPLNNPLPRERTPFALQTINALEEESRGFPTQVERIFQESVYKTQQTIWKETRNMVQRQFIA tlr4_human Toll-like receptor 4 TLR4 hToll protein protein human Homo sapiens Human MMSASRLAGTLIPAMAFLSCVRPESWEPCVEVVPNITYQCMELNFYKIPDNLPFSTKNLDLSFNPLRHLGSYSFFSFPELQVLDLSRCEIQTIEDGAYQSLSHLSTLILTGNPIQSLALGAFSGLSSLQKLVAVETNLASLENFPIGHLKTLKELNVAHNLIQSFKLPEYFSNLTNLEHLDLSSNKIQSIYCTDLRVLHQMPLLNLSLDLSLNPMNFIQPGAFKEIRLHKLTLRNNFDSLNVMKTCIQGLAGLEVHRLVLGEFRNEGNLEKFDKSALEGLCNLTIEEFRLAYLDYYLDDIIDLFNCLTNVSSFSLVSVTIERVKDFSYNFGWQHLELVNCKFGQFPTLKLKSLKRLTFTSNKGGNAFSEVDLPSLEFLDLSRNGLSFKGCCSQSDFGTTSLKYLDLSFNGVITMSSNFLGLEQLEHLDFQHSNLKQMSEFSVFLSLRNLIYLDISHTHTRVAFNGIFNGLSSLEVLKMAGNSFQENFLPDIFTELRNLTFLDLSQCQLEQLSPTAFNSLSSLQVLNMSHNNFFSLDTFPYKCLNSLQVLDYSLNHIMTSKKQELQHFPSSLAFLNLTQNDFACTCEHQSFLQWIKDQRQLLVEVERMECATPSDKQGMPVLSLNITCQMNKTIIGVSVLSVLVVSVVAVLVYKFYFHLMLLAGCIKYGRGENIYDAFVIYSSQDEDWVRNELVKNLEEGVPPFQLCLHYRDFIPGVAIAANIIHEGFHKSRKVIVVVSQHFIQSRWCIFEYEIAQTWQFLSSRAGIIFIVLQKVEKTLLRQQVELYRLLSRNTYLEWEDSVLGRHIFWRRLRKALLDGKSWNPEGTVGTGCNWQEATSI ager-tirap-1 3435515 N/A 3435521 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell engineered engineered myc tag 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 ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false N/A 3435522 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell engineered engineered His tag 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 ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false cytosolic domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Fig. S2A S100A11 ager-tirap-2 3435515 N/A 3435523 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell engineered engineered myc tag 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 ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false N/A 3435521 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell engineered engineered myc tag 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 ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false N/A 3435522 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell engineered engineered His tag 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 ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false cytosolic domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Q15109:p.Ser391Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false Q15109:p.Ser391Glu mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain false association association false false false Fig. S2B ager-myd88-3 3435515 N/A 3435523 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell engineered engineered myc tag 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 ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false N/A 3435521 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell engineered engineered myc tag 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 ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false P58753:p.Leu118_Ser131delinsSerGlnSerSerSerAlaThrGlyGlyGlyAlaSerGlySer mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false N/A 3435522 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell engineered engineered His tag 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 ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false cytosolic domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false association association false false false Fig. S2B ager-myd88-4 3435515 N/A 3435523 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell engineered engineered Q99836:p.Arg196_Thr202delinsSerSerGlyLeuGlyGlyThr mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false myc tag 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 ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false N/A 3435521 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell engineered engineered myc tag 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 ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false N/A 3435522 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell engineered engineered His tag 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 ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false cytosolic domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false association association false false false Fig. S2B ager-tirap-3 3435515 N/A 3435524 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell engineered engineered ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false N/A 3435522 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell engineered engineered His tag 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 ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false cytosolic domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Q15109:p.Ser391Glu mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false Q15109:p.Ser391Ala mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Fig. S1G binding of PKCζ to RAGE was independent of the phosphorylation status (Figure 1G and S1G). rage-pkc-1 3435516 N/A 3435524 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3435522 anti tag western anti tag western blot anti tag western unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell engineered engineered His tag 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 Q15109:p.Ser391Ala mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false phosphorylated Ser opser O-phospho-L-serine (2S)-2-amino-3-(phosphonooxy)propanoic acid 2-amino-3-hydroxypropanoic acid 3-phosphate 2-azanyl-3-(phosphonooxy)propanoic acid ACT_SITE Phosphoserine intermediate MOD_RES Phosphoserine O-phospho-L-serine O-phosphonoserine O-phosphorylated L-serine O3-phosphoserine serine phosphate ester OPSer Phospho Phospho Seryl Phosphorylation phosphorylations mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false physical association physical association false false false Fig. 1G S100A11 ager-prkcz-1 3435516 N/A 3435521 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3435523 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3435524 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3435522 anti tag western anti tag western blot anti tag western unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell engineered engineered 32p radiolabel 32p radiolabel 32p radiolabel P32 32P 32P P32 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false His tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false cytosolic domain required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain false ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false myc tag myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false flag tag flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term range false association association false false false Fig. 2A S100A11 S100A12 HMGB1 RAGE TIRAP and MyD88, but not TRAM, were co-precipitated with overexpressed RAGE and RAGE was phosphorylated in HEK293 cells predominantly when the cells were treated with RAGE ligands, S100A11, S100A12, HMGB1 and AGE but not with a TLR4 ligand (LPS) and TLR2 ligands (Figure 2A). ager-prkcz-2 3435516 N/A 3435525 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3435521 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3435523 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3435522 anti tag western anti tag western blot anti tag western unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell engineered engineered His tag 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 Q15109:p.Ser391Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false phosphorylated Ser opser O-phospho-L-serine (2S)-2-amino-3-(phosphonooxy)propanoic acid 2-amino-3-hydroxypropanoic acid 3-phosphate 2-azanyl-3-(phosphonooxy)propanoic acid ACT_SITE Phosphoserine intermediate MOD_RES Phosphoserine O-phospho-L-serine O-phosphonoserine O-phosphorylated L-serine O3-phosphoserine serine phosphate ester OPSer Phospho Phospho Seryl Phosphorylation phosphorylations mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false association association false false false Fig. 2B AGE AGE (100 µg/ml)-induced phosphorylation of RAGE at Ser391 is essential for recruitment of endogenous TIRAP and MyD88 (B: The results were confirmed by an independent and two related experiments.) and downstream signal transduction (C: The results were confirmed by an independent and two related experiments.) in HEK293 cells. ager-myd88-2 3435516 N/A 3435524 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3435522 anti tag western anti tag western blot anti tag western unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell engineered engineered 32p radiolabel 32p radiolabel 32p radiolabel P32 32P 32P P32 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false His tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false cytosolic domain required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain false ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false myc tag myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false flag tag flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged c-term range C-terminal range c-term range c-term range C-terminal range c-term range false physical association physical association false false false Fig. 2A RAGE interacts exclusively with PKCzeta when HUVECs are treated with LPS and TLR2. rage-1 3435517 N/A 3435526 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3435527 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3435523 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell myc tag 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 ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false N/A 3435528 unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3435521 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell myc tag 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 ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false association association false false false Fig. 2D Essential nature of TIRAP and MyD88 for RAGE signaling was demonstrated using TIRAP-down regulated NIH3T3 cells and MyD88-/- mouse fibroblasts (Figures S2D, 2D and 2E). Overexpression of TIRAP or MyD88 rescued the defective protein interaction and signal transduction in corresponding cells. These results indicate that TIRAP and MyD88 function as essential adaptor proteins for RAGE, binding to ligand-activated phosphorylated RAGE and transducing a signal from it. AGE ager-irak4-2 3435517 N/A 3435527 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3435528 unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3435521 anti tag western anti tag western blot anti tag western unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell myc tag 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 ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false association association false false false Fig. 2D Essential nature of TIRAP and MyD88 for RAGE signaling was demonstrated using TIRAP-down regulated NIH3T3 cells and MyD88-/- mouse fibroblasts (Figures S2D, 2D and 2E). Overexpression of TIRAP or MyD88 rescued the defective protein interaction and signal transduction in corresponding cells. These results indicate that TIRAP and MyD88 function as essential adaptor proteins for RAGE, binding to ligand-activated phosphorylated RAGE and transducing a signal from it. AGE ager-irak4-3 3435517 N/A 3435526 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3435529 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3435527 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3435523 anti tag western anti tag western blot anti tag western unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell myc tag 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 ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false N/A 3435528 unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level association association false false false Fig. 2D Essential nature of TIRAP and MyD88 for RAGE signaling was demonstrated using TIRAP-down regulated NIH3T3 cells and MyD88-/- mouse fibroblasts (Figures S2D, 2D and 2E). Overexpression of TIRAP or MyD88 rescued the defective protein interaction and signal transduction in corresponding cells. These results indicate that TIRAP and MyD88 function as essential adaptor proteins for RAGE, binding to ligand-activated phosphorylated RAGE and transducing a signal from it. AGE ager-prkcz-3 3435517 N/A 3435527 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3435528 unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association false false false Fig. 2D Essential nature of TIRAP and MyD88 for RAGE signaling was demonstrated using TIRAP-down regulated NIH3T3 cells and MyD88-/- mouse fibroblasts (Figures S2D, 2D and 2E). Overexpression of TIRAP or MyD88 rescued the defective protein interaction and signal transduction in corresponding cells. These results indicate that TIRAP and MyD88 function as essential adaptor proteins for RAGE, binding to ligand-activated phosphorylated RAGE and transducing a signal from it. ager-irak4-4 3435518 N/A 3435526 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3435530 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3435527 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3435521 anti tag western anti tag western blot anti tag western unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell myc tag 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 ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false N/A 3435528 unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level association association false false false Fig. 2D Essential nature of TIRAP and MyD88 for RAGE signaling was demonstrated using TIRAP-down regulated NIH3T3 cells and MyD88-/- mouse fibroblasts (Figures S2D, 2D and 2E). Overexpression of TIRAP or MyD88 rescued the defective protein interaction and signal transduction in corresponding cells. These results indicate that TIRAP and MyD88 function as essential adaptor proteins for RAGE, binding to ligand-activated phosphorylated RAGE and transducing a signal from it. AGE ager-prkcz-5 3435518 N/A 3435527 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3435528 unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association false false false Fig. 2D Essential nature of TIRAP and MyD88 for RAGE signaling was demonstrated using TIRAP-down regulated NIH3T3 cells and MyD88-/- mouse fibroblasts (Figures S2D, 2D and 2E). Overexpression of TIRAP or MyD88 rescued the defective protein interaction and signal transduction in corresponding cells. These results indicate that TIRAP and MyD88 function as essential adaptor proteins for RAGE, binding to ligand-activated phosphorylated RAGE and transducing a signal from it. ager-irak4-5 3435518 N/A 3435526 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3435527 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3435523 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell myc tag 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 ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false N/A 3435528 unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3435521 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell myc tag 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 ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false association association false false false Fig. 2D Essential nature of TIRAP and MyD88 for RAGE signaling was demonstrated using TIRAP-down regulated NIH3T3 cells and MyD88-/- mouse fibroblasts (Figures S2D, 2D and 2E). Overexpression of TIRAP or MyD88 rescued the defective protein interaction and signal transduction in corresponding cells. These results indicate that TIRAP and MyD88 function as essential adaptor proteins for RAGE, binding to ligand-activated phosphorylated RAGE and transducing a signal from it. AGE ager-irak4-6 3435519 N/A 3435531 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3435525 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3435521 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3435523 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3435522 anti tag western anti tag western blot anti tag western unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell engineered engineered His tag 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 ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false association association false false false Fig. 4A, Fig. S5B RAGE was co-precipitated with TIRAP, MyD88, TRAM and IRAK-4 when the receptor was activated by treatment with LBP-LPS, HMGB1-LPS and S100A8/Ap (weakly) ager-irak4-7 3435519 N/A 3435525 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3435521 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3435523 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3435522 anti tag western anti tag western blot anti tag western unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell engineered engineered His tag 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 ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false association association false false false Fig. 4A, Fig. S5B RAGE was co-precipitated with TIRAP, MyD88 and IRAK-4 when the receptors were activated by treatment with AGE, HMGB1-LPS and S100A8/Ap (weakly) ager-irak4-8 3435519 N/A 3435532 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell engineered engineered ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false myc tag 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 3435531 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3435525 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3435521 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3435523 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level association association false false false Fig. 4A, Fig. S5B TLR4 was co-precipitated with TRAM in addition to the three proteins only when the receptors were activated by the ligands (Figure 4A and S5B). ager-myd88-1 3435520 N/A 3435525 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3435521 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3435523 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3435524 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3435522 unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level association association false false false Fig. 3A, Fig. S3 Application to HUVECs with RAGE ligands such as S100A11, S100A12, HMGB1, and AGE but not LPS and TLR2 ligands induced phosphorylation of endogenous RAGE, interaction with TIRAP, MyD88, and IRAK4, and transduction of signal to the downstream molecules (Figures 3A and 3B). S100A11 S100A12 HMGB1 AGE ager-irak4-1 3435520 N/A 3435522 unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3435524 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association false false false Fig. 3A, Fig. S3 RAGE interacts exclusively with PKCzeta when HUVECs are treated with LPS and TLR2.