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/ TIRAP, an adaptor protein for TLR2/4, transduces a signal from RAGE phosphorylated upon ligand binding. TIRAP, an adaptor protein for TLR2/4, transduces a signal from RAGE phosphorylated upon ligand binding. PloS one 2010 imex curation Sakaguchi M., Murata H., Yamamoto K., Ono T., Sakaguchi Y., Motoyama A., Hibino T., Kataoka K., Huh NH. in SFig1G PKCzeta is human as provided by authors. I haven't annotated any of the kinase assays in Fig 1 and Fig. S1 as RAGE was not highly purified, but only immunopurified. Suppl Fig 5C and Fig 4B are not in since ip is carried out using anti-myc in cells transfected with two proteins with myc-tag. namu@md.okayama-u.ac.jp 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation 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. in SFig1G PKCzeta is human as provided by authors. I haven't annotated any of the kinase assays in Fig 1 and Fig. S1 as RAGE was not highly purified, but only immunopurified. Suppl Fig 5C and Fig 4B are not in since ip is carried out using anti-myc in cells transfected with two proteins with myc-tag. namu@md.okayama-u.ac.jp Accepted 2018-JUN-04 AT 13:46 BST AT 13:46 BST by ORCHARD PloS one 2010 imex curation True, I've removed tag and modified exp-prep accordingly; I prefer not to curate them since the ip is in vivo and the molecular weight is not ok in these conditions; removed the 2-baits interactions and added those with only anti-myc TIRAP, an adaptor protein for TLR2/4, transduces a signal from RAGE phosphorylated upon ligand binding. TIRAP, an adaptor protein for TLR2/4, transduces a signal from RAGE phosphorylated upon ligand binding. PloS one 2010 imex curation Sakaguchi M., Murata H., Yamamoto K., Ono T., Sakaguchi Y., Motoyama A., Hibino T., Kataoka K., Huh NH. in SFig1G PKCzeta is human as provided by authors. I haven't annotated any of the kinase assays in Fig 1 and Fig. S1 as RAGE was not highly purified, but only immunopurified. Suppl Fig 5C and Fig 4B are not in since ip is carried out using anti-myc in cells transfected with two proteins with myc-tag. namu@md.okayama-u.ac.jp 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation 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. in SFig1G PKCzeta is human as provided by authors. I haven't annotated any of the kinase assays in Fig 1 and Fig. S1 as RAGE was not highly purified, but only immunopurified. Suppl Fig 5C and Fig 4B are not in since ip is carried out using anti-myc in cells transfected with two proteins with myc-tag. 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 Accepted 2018-JUN-04 AT 13:46 BST AT 13:46 BST by ORCHARD PloS one 2010 imex curation True, I've removed tag and modified exp-prep accordingly; I prefer not to curate them since the ip is in vivo and the molecular weight is not ok in these conditions; removed the 2-baits interactions and added those with only anti-myc TIRAP, an adaptor protein for TLR2/4, transduces a signal from RAGE phosphorylated upon ligand binding. TIRAP, an adaptor protein for TLR2/4, transduces a signal from RAGE phosphorylated upon ligand binding. PloS one 2010 imex curation Sakaguchi M., Murata H., Yamamoto K., Ono T., Sakaguchi Y., Motoyama A., Hibino T., Kataoka K., Huh NH. in SFig1G PKCzeta is human as provided by authors. I haven't annotated any of the kinase assays in Fig 1 and Fig. S1 as RAGE was not highly purified, but only immunopurified. Suppl Fig 5C and Fig 4B are not in since ip is carried out using anti-myc in cells transfected with two proteins with myc-tag. namu@md.okayama-u.ac.jp 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation mouse-fibroblast Mus musculus fibroblast Fibroblast 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. in SFig1G PKCzeta is human as provided by authors. I haven't annotated any of the kinase assays in Fig 1 and Fig. S1 as RAGE was not highly purified, but only immunopurified. Suppl Fig 5C and Fig 4B are not in since ip is carried out using anti-myc in cells transfected with two proteins with myc-tag. 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 Accepted 2018-JUN-04 AT 13:46 BST AT 13:46 BST by ORCHARD PloS one 2010 imex curation True, I've removed tag and modified exp-prep accordingly; I prefer not to curate them since the ip is in vivo and the molecular weight is not ok in these conditions; removed the 2-baits interactions and added those with only anti-myc TIRAP, an adaptor protein for TLR2/4, transduces a signal from RAGE phosphorylated upon ligand binding. TIRAP, an adaptor protein for TLR2/4, transduces a signal from RAGE phosphorylated upon ligand binding. PloS one 2010 imex curation Sakaguchi M., Murata H., Yamamoto K., Ono T., Sakaguchi Y., Motoyama A., Hibino T., Kataoka K., Huh NH. in SFig1G PKCzeta is human as provided by authors. I haven't annotated any of the kinase assays in Fig 1 and Fig. S1 as RAGE was not highly purified, but only immunopurified. Suppl Fig 5C and Fig 4B are not in since ip is carried out using anti-myc in cells transfected with two proteins with myc-tag. namu@md.okayama-u.ac.jp 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation 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. in SFig1G PKCzeta is human as provided by authors. I haven't annotated any of the kinase assays in Fig 1 and Fig. S1 as RAGE was not highly purified, but only immunopurified. Suppl Fig 5C and Fig 4B are not in since ip is carried out using anti-myc in cells transfected with two proteins with myc-tag. namu@md.okayama-u.ac.jp Accepted 2018-JUN-04 AT 13:46 BST AT 13:46 BST by ORCHARD PloS one 2010 imex curation True, I've removed tag and modified exp-prep accordingly; I prefer not to curate them since the ip is in vivo and the molecular weight is not ok in these conditions; removed the 2-baits interactions and added those with only anti-myc TIRAP, an adaptor protein for TLR2/4, transduces a signal from RAGE phosphorylated upon ligand binding. TIRAP, an adaptor protein for TLR2/4, transduces a signal from RAGE phosphorylated upon ligand binding. PloS one 2010 imex curation Sakaguchi M., Murata H., Yamamoto K., Ono T., Sakaguchi Y., Motoyama A., Hibino T., Kataoka K., Huh NH. in SFig1G PKCzeta is human as provided by authors. I haven't annotated any of the kinase assays in Fig 1 and Fig. S1 as RAGE was not highly purified, but only immunopurified. Suppl Fig 5C and Fig 4B are not in since ip is carried out using anti-myc in cells transfected with two proteins with myc-tag. namu@md.okayama-u.ac.jp 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation 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. in SFig1G PKCzeta is human as provided by authors. I haven't annotated any of the kinase assays in Fig 1 and Fig. S1 as RAGE was not highly purified, but only immunopurified. Suppl Fig 5C and Fig 4B are not in since ip is carried out using anti-myc in cells transfected with two proteins with myc-tag. 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 Accepted 2018-JUN-04 AT 13:46 BST AT 13:46 BST by ORCHARD PloS one 2010 imex curation True, I've removed tag and modified exp-prep accordingly; I prefer not to curate them since the ip is in vivo and the molecular weight is not ok in these conditions; removed the 2-baits interactions and added those with only anti-myc TIRAP, an adaptor protein for TLR2/4, transduces a signal from RAGE phosphorylated upon ligand binding. TIRAP, an adaptor protein for TLR2/4, transduces a signal from RAGE phosphorylated upon ligand binding. PloS one 2010 imex curation Sakaguchi M., Murata H., Yamamoto K., Ono T., Sakaguchi Y., Motoyama A., Hibino T., Kataoka K., Huh NH. in SFig1G PKCzeta is human as provided by authors. I haven't annotated any of the kinase assays in Fig 1 and Fig. S1 as RAGE was not highly purified, but only immunopurified. Suppl Fig 5C and Fig 4B are not in since ip is carried out using anti-myc in cells transfected with two proteins with myc-tag. namu@md.okayama-u.ac.jp 09-May-2019: Contacted by IntAct-Help. Only protein-protein interactions imex curation 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. in SFig1G PKCzeta is human as provided by authors. I haven't annotated any of the kinase assays in Fig 1 and Fig. S1 as RAGE was not highly purified, but only immunopurified. Suppl Fig 5C and Fig 4B are not in since ip is carried out using anti-myc in cells transfected with two proteins with myc-tag. 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 Accepted 2018-JUN-04 AT 13:46 BST AT 13:46 BST by ORCHARD PloS one 2010 imex curation True, I've removed tag and modified exp-prep accordingly; I prefer not to curate them since the ip is in vivo and the molecular weight is not ok in these conditions; removed the 2-baits interactions and added those with only anti-myc myd88_human Myeloid differentiation primary response protein MyD88 MYD88 protein protein human Homo sapiens Human MAAGGPGAGSAAPVSSTSSLPLAALNMRVRRRLSLFLNVRTQVAADWTALAEEMDFEYLEIRQLETQADPTGRLLDAWQGRPGASVGRLLELLTKLGRDDVLLELGPSIEEDCQKYILKQQQEEAEKPLQVAAVDSSVPRTAELAGITTLDDPLGHMPERFDAFICYCPSDIQFVQEMIRQLEQTNYRLKLCVSDRDVLPGTCVWSIASELIEKRCRRMVVVVSDDYLQSKECDFQTKFALSLSPGAHQKRLIPIKYKAMKKEFPSILRFITVCDYTNPCTKSWFWTRLAKALSLP CEAE3F6B99524333 kpcz_human Protein kinase C zeta type PRKCZ PKC2 nPKC-zeta protein protein human Homo sapiens Human MPSRTGPKMEGSGGRVRLKAHYGGDIFITSVDAATTFEELCEEVRDMCRLHQQHPLTLKWVDSEGDPCTVSSQMELEEAFRLARQCRDEGLIIHVFPSTPEQPGLPCPGEDKSIYRRGARRWRKLYRANGHLFQAKRFNRRAYCGQCSERIWGLARQGYRCINCKLLVHKRCHGLVPLTCRKHMDSVMPSQEPPVDDKNEDADLPSEETDGIAYISSSRKHDSIKDDSEDLKPVIDGMDGIKISQGLGLQDFDLIRVIGRGSYAKVLLVRLKKNDQIYAMKVVKKELVHDDEDIDWVQTEKHVFEQASSNPFLVGLHSCFQTTSRLFLVIEYVNGGDLMFHMQRQRKLPEEHARFYAAEICIALNFLHERGIIYRDLKLDNVLLDADGHIKLTDYGMCKEGLGPGDTTSTFCGTPNYIAPEILRGEEYGFSVDWWALGVLMFEMMAGRSPFDIITDNPDMNTEDYLFQVILEKPIRIPRFLSVKASHVLKGFLNKDPKERLGCRPQTGFSDIKSHAFFRSIDWDLLEKKQALPPFQPQITDDYGLDNFDTQFTSEPVQLTPDDEDAIKRIDQSEFEGFEYINPLLLSTEESV 88CCD0577E6A596C 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 C387D20229E12265 kpcz_mouse Protein kinase C zeta type nPKC-zeta Prkcz Pkcz protein protein mouse Mus musculus Mouse MPSRTDPKMDRSGGRVRLKAHYGGDILITSVDAMTTFKDLCEEVRDMCGLHQQHPLTLKWVDSEGDPCTVSSQMELEEAFRLVCQGRDEVLIIHVFPSIPEQPGMPCPGEDKSIYRRGARRWRKLYRANGHLFQAKRFNRGAYCGQCSERIWGLSRQGYRCINCKLLVHKRCHVLVPLTCRRHMDSVMPSQEPPVDDKNDGVDLPSEETDGIAYISSSRKHDNIKDDSEDLKPVIDGVDGIKISQGLGLQDFDLIRVIGRGSYAKVLLVRLKKNDQIYAMKVVKKELVHDDEDIDWVQTEKHVFEQASSNPFLVGLHSCFQTTSRLFLVIEYVNGGDLMFHMQRQRKLPEEHARFYAAEICIALNFLHERGIIYRDLKLDNVLLDADGHIKLTDYGMCKEGLGPGDTTSTFCGTPNYIAPEILRGEEYGFSVDWWALGVLMFEMMAGRSPFDIITDNPDMNTEDYLFQVILEKPIRIPRFLSVKASHVLKGFLNKDPKERLGCRPQTGFSDIKSHAFFRSIDWDLLEKKQTLPPFQPQITDDYGLDNFDTQFTSEPVQLTPDDEDVIKRIDQSEFEGFEYINPLLLSAEESV 690AD891C25BC311 rage_mouse Advanced glycosylation end product-specific receptor Ager Rage Receptor for advanced glycosylation end products protein protein mouse Mus musculus Mouse MPAGTAARAWVLVLALWGAVAGGQNITARIGEPLVLSCKGAPKKPPQQLEWKLNTGRTEAWKVLSPQGGPWDSVARILPNGSLLLPATGIVDEGTFRCRATNRRGKEVKSNYRVRVYQIPGKPEIVDPASELTASVPNKVGTCVSEGSYPAGTLSWHLDGKLLIPDGKETLVKEETRRHPETGLFTLRSELTVIPTQGGTHPTFSCSFSLGLPRRRPLNTAPIQLRVREPGPPEGIQLLVEPEGGIVAPGGTVTLTCAISAQPPPQVHWIKDGAPLPLAPSPVLLLPEVGHEDEGTYSCVATHPSHGPQESPPVSIRVTETGDEGPAEGSVGESGLGTLALALGILGGLGVVALLVGAILWRKRQPRREERKAPESQEDEEERAELNQSEEAEMPENGAGGP DBFDC50A6C8CB902 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 DIP protein P58753 original sequence version: 119 44A8454D04704540 rage_human Advanced glycosylation end product-specific receptor AGER RAGE Receptor for advanced glycosylation end products protein protein human Homo sapiens Human MAAGTAVGAWVLVLSLWGAVVGAQNITARIGEPLVLKCKGAPKKPPQRLEWKLNTGRTEAWKVLSPQGGGPWDSVARVLPNGSLFLPAVGIQDEGIFRCQAMNRNGKETKSNYRVRVYQIPGKPEIVDSASELTAGVPNKVGTCVSEGSYPAGTLSWHLDGKPLVPNEKGVSVKEQTRRHPETGLFTLQSELMVTPARGGDPRPTFSCSFSPGLPRHRALRTAPIQPRVWEPVPLEEVQLVVEPEGGAVAPGGTVTLTCEVPAQPSPQIHWMKDGVPLPLPPSPVLILPEIGPQDQGTYSCVATHSSHGPQESRAVSISIIEPGEEGPTAGSVGGSGLGTLALALGILGGLGTAALLIGVILWQRRQRRGEERKAPENQEEEEERAELNQSEEPEAGESSTGGP DIP protein Q15109 original sequence version: 157 0D584C436C30CCE7 irak4_mouse Interleukin-1 receptor-associated kinase 4 Irak4 protein protein mouse Mus musculus Mouse MNKPLTPSTYIRNLNVGILRKLSDFIDPQEGWKKLAVAIKKPSGDDRYNQFHIRRFEALLQTGKSPTCELLFDWGTTNCTVGDLVDLLVQIELFAPATLLLPDAVPQTVKSLPPREAATVAQTHGPCQEKDRTSVMPMPKLEHSCEPPDSSSPDNRSVESSDTRFHSFSFHELKSITNNFDEQPASAGGNRMGEGGFGVVYKGCVNNTIVAVKKLGAMVEISTEELKQQFDQEIKVMATCQHENLVELLGFSSDSDNLCLVYAYMPNGSLLDRLSCLDGTPPLSWHTRCKVAQGTANGIRFLHENHHIHRDIKSANILLDKDFTAKISDFGLARASARLAQTVMTSRIVGTTAYMAPEALRGEITPKSDIYSFGVVLLELITGLAAVDENREPQLLLDIKEEIEDEEKTIEDYTDEKMSDADPASVEAMYSAASQCLHEKKNRRPDIAKVQQLLQEMSA DIP protein Q8R4K2 original sequence version: 107 DIP protein Q8R4K2 original sequence version: 98 DIP protein Q8R4K2 original sequence version: 77 FC11AD06983B7AEB myd88_mouse Myeloid differentiation primary response protein MyD88 Myd88 protein protein mouse Mus musculus Mouse MSAGDPRVGSGSLDSFMFSIPLVALNVGVRRRLSLFLNPRTPVAADWTLLAEEMGFEYLEIRELETRPDPTRSLLDAWQGRSGASVGRLLELLALLDREDILKELKSRIEEDCQKYLGKQQNQESEKPLQVARVESSVPQTKELGGITTLDDPLGQTPELFDAFICYCPNDIEFVQEMIRQLEQTDYRLKLCVSDRDVLPGTCVWSIASELIEKRCRRMVVVVSDDYLQSKECDFQTKFALSLSPGVQQKRLIPIKYKAMKKDFPSILRFITICDYTNPCTKSWFWTRLAKALSLP DIP protein P22366 original sequence version: 131 DIP protein P22366 original sequence version: 141 DIP protein P22366-1 original sequence version: 143 DIP protein P22366-1 original sequence version: 144 DIP protein P22366 original sequence version: 122 393A75C3723FFE69 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 2D1171F9B04D9C4C irak4_human Interleukin-1 receptor-associated kinase 4 Renal carcinoma antigen NY-REN-64 IRAK4 protein protein human Homo sapiens Human MNKPITPSTYVRCLNVGLIRKLSDFIDPQEGWKKLAVAIKKPSGDDRYNQFHIRRFEALLQTGKSPTSELLFDWGTTNCTVGDLVDLLIQNEFFAPASLLLPDAVPKTANTLPSKEAITVQQKQMPFCDKDRTLMTPVQNLEQSYMPPDSSSPENKSLEVSDTRFHSFSFYELKNVTNNFDERPISVGGNKMGEGGFGVVYKGYVNNTTVAVKKLAAMVDITTEELKQQFDQEIKVMAKCQHENLVELLGFSSDGDDLCLVYVYMPNGSLLDRLSCLDGTPPLSWHMRCKIAQGAANGINFLHENHHIHRDIKSANILLDEAFTAKISDFGLARASEKFAQTVMTSRIVGTTAYMAPEALRGEITPKSDIYSFGVVLLEIITGLPAVDEHREPQLLLDIKEEIEDEEKTIEDYIDKKMNDADSTSVEAMYSVASQCLHEKKNKRPDIKKVQQLLQEMTAS DIP protein Q9NWZ3 original sequence version: 93 DIP protein Q9NWZ3 original sequence version: 92 DIP protein Q9NWZ3 original sequence version: 144 6C8156ADF25FF81E tlr4_human Toll-like receptor 4 TLR4 hToll protein protein human Homo sapiens Human MMSASRLAGTLIPAMAFLSCVRPESWEPCVEVVPNITYQCMELNFYKIPDNLPFSTKNLDLSFNPLRHLGSYSFFSFPELQVLDLSRCEIQTIEDGAYQSLSHLSTLILTGNPIQSLALGAFSGLSSLQKLVAVETNLASLENFPIGHLKTLKELNVAHNLIQSFKLPEYFSNLTNLEHLDLSSNKIQSIYCTDLRVLHQMPLLNLSLDLSLNPMNFIQPGAFKEIRLHKLTLRNNFDSLNVMKTCIQGLAGLEVHRLVLGEFRNEGNLEKFDKSALEGLCNLTIEEFRLAYLDYYLDDIIDLFNCLTNVSSFSLVSVTIERVKDFSYNFGWQHLELVNCKFGQFPTLKLKSLKRLTFTSNKGGNAFSEVDLPSLEFLDLSRNGLSFKGCCSQSDFGTTSLKYLDLSFNGVITMSSNFLGLEQLEHLDFQHSNLKQMSEFSVFLSLRNLIYLDISHTHTRVAFNGIFNGLSSLEVLKMAGNSFQENFLPDIFTELRNLTFLDLSQCQLEQLSPTAFNSLSSLQVLNMSHNNFFSLDTFPYKCLNSLQVLDYSLNHIMTSKKQELQHFPSSLAFLNLTQNDFACTCEHQSFLQWIKDQRQLLVEVERMECATPSDKQGMPVLSLNITCQMNKTIIGVSVLSVLVVSVVAVLVYKFYFHLMLLAGCIKYGRGENIYDAFVIYSSQDEDWVRNELVKNLEEGVPPFQLCLHYRDFIPGVAIAANIIHEGFHKSRKVIVVVSQHFIQSRWCIFEYEIAQTWQFLSSRAGIIFIVLQKVEKTLLRQQVELYRLLSRNTYLEWEDSVLGRHIFWRRLRKALLDGKSWNPEGTVGTGCNWQEATSI 92C48F55821133E8 rage-pkc-1 4 8 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 13 anti tag western anti tag western blot anti tag western western blot western blot Immuno blot Immuno blot 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 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 S A Tue Jun 30 19:46:23 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser391Ala' 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 autoradiography autoradiography certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm 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 physical association physical association Fig. 1G S100A11 ager-prkcz-1 4 12 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 7 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 8 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 13 anti tag western anti tag western blot anti tag western western blot western blot Immuno blot Immuno blot 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 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 cytosolic domain required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain 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 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 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 association association 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 4 17 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 12 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 7 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 13 anti tag western anti tag western blot anti tag western western blot western blot Immuno blot Immuno blot 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 Q15109:p.Ser391Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain S A Tue Jun 30 19:59:57 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser391Ala' 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 autoradiography autoradiography certain certain sequence position certain certain certain sequence position certain prerequisite-ptm prerequisite-ptm prerequisite ptm required ptm prerequisite-ptm required-ptm 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 association association 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. 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-myd88-1 1 17 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 12 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 7 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 8 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 13 unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level association association 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 1 13 unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 8 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association Fig. 3A, Fig. S3 RAGE interacts exclusively with PKCzeta when HUVECs are treated with LPS and TLR2. ager-myd88-2 4 8 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 13 anti tag western anti tag western blot anti tag western western blot western blot Immuno blot Immuno blot 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 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 cytosolic domain required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain 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 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 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 physical association physical association Fig. 2A RAGE interacts exclusively with PKCzeta when HUVECs are treated with LPS and TLR2. rage-1 3 14 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 10 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 7 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 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 11 unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 12 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 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 association association 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 3 10 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 11 unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 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 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 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 association association 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 3 14 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 9 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 10 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 7 anti tag western anti tag western blot anti tag western western blot western blot Immuno blot Immuno blot 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 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 11 unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level association association 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 3 10 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 11 unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association 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 5 14 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 15 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 10 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 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 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 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 11 unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level association association 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 5 10 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 11 unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association 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 5 14 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 10 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 7 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 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 11 unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 12 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 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 association association 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-tirap-1 6 12 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 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 13 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 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 cytosolic domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association Fig. S2A S100A11 ager-tirap-2 6 7 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 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 12 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 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 13 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 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 cytosolic domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain Q15109:p.Ser391Ala mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain S A Tue Jun 30 17:39:59 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser391Ala' Q15109:p.Ser391Glu mutation increasing strength mutation increasing interaction strength mutation increasing strength certain certain sequence position certain certain certain sequence position certain S E Tue Jun 30 17:32:30 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser391Glu' association association Fig. S2B ager-myd88-3 6 7 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 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 12 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 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 P58753:p.Leu118_Ser131delinsSerGlnSerSerSerAlaThrGlyGlyGlyAlaSerGlySer mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain LQLRDATPGGAIVS SQSSSATGGGASGS Wed Jun 13 17:57:42 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Leu118_Ser13delinsSerGlnSerSerSerAlaThrGlyGlyGlyAlaSerGlySer' Tue Jun 30 19:42:20 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Leu118_Ser131delinsSerGlnSerSerSerAlaThrGlyGlyGlyAlaSerGlySer' 13 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 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 cytosolic domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain association association Fig. S2B I am not sure these interaction is suitable for being captured. Dear reviewer, if you think this interaction has to be deleted, please delete it ;-) ager-myd88-4 6 7 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 RDVLPGT SSGLGGT Tue Jun 30 17:36:59 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg196_Thr202delinsSerSerGlyLeuGlyGlyThr' 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 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 12 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 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 13 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 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 cytosolic domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain association association Fig. S2B I am not sure these interaction is suitable for being captured. Dear reviewer, if you think this interaction has to be deleted, please delete it ;-) ager-irak4-6 2 16 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 17 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 12 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 7 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 13 anti tag western anti tag western blot anti tag western western blot western blot Immuno blot Immuno blot 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 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 association association Fig. 4A, Fig. S5B Immunoprecipitation using antibodies against respective tags of TLR4 and RAGE followed by Western blot analysis showed that RAGE was co-precipitated with TIRAP, MyD88 and IRAK-4, while 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). 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 2 17 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 12 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 7 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 13 anti tag western anti tag western blot anti tag western western blot western blot Immuno blot Immuno blot 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 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 association association 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-tirap-3 6 8 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 13 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 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 cytosolic domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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 S E Tue Jun 30 18:05:46 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser391Glu' 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 S A Tue Jun 30 16:23:31 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser391Ala' physical association physical association Fig. S1G binding of PKCζ to RAGE was independent of the phosphorylation status (Figure 1G and S1G). binding of PKCζ to RAGE was independent of the phosphorylation status (Figure 1G and S1G). ager-irak4-8 2 18 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 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 16 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 17 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 12 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level 7 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level association association Fig. 4A, Fig. S5B Immunoprecipitation using antibodies against respective tags of TLR4 and RAGE followed by Western blot analysis showed that RAGE was co-precipitated with TIRAP, MyD88 and IRAK-4, while 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). 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).