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/ The functional effects of physical interactions among Toll-like receptors 7, 8, and 9. The functional effects of physical interactions among Toll-like receptors 7, 8, and 9. The Journal of biological chemistry 2006 imex curation Wang J., Shao Y., Bennett TA., Shankar RA., Wightman PD., Reddy LG. The (NCBI) Human TLR8 "isoform 1" (1059 aa-NP_057694.2 ) used here, corresponds to UniProt TLR8 isoform 2. lgreddy@mmm.com 27-Mar-2018: 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 Wang J., Shao Y., Bennett TA., Shankar RA., Wightman PD., Reddy LG. The (NCBI) Human TLR8 "isoform 1" (1059 aa-NP_057694.2 ) used here, corresponds to UniProt TLR8 isoform 2. lgreddy@mmm.com Accepted 2018-MAR-15 AT 08:35 GMT AT 08:35 GMT by LIVIAP The Journal of biological chemistry 2006 Only protein-protein interactions imex curation imex curation The HA tag is not at the "N-terminUS" - it is "n-terminAL" according to text :- "The TLR8 expression vector with N-terminal HA tag was constructed by inserting the HA tag at the C-terminal end of the predicted signal peptide sequence (after 45 residues from the N-terminal end or TLR8 isoform 1)". I think they mean "OF TLR8 isoform 1" not "or isoform 1". I described the tag as "N-terminal" as they do in the text, as opposed to "N-terminus" which I would use if the tag was attached, as usual, at aas 1-~3 of a sequence. The functional effects of physical interactions among Toll-like receptors 7, 8, and 9. The functional effects of physical interactions among Toll-like receptors 7, 8, and 9. The Journal of biological chemistry 2006 imex curation Wang J., Shao Y., Bennett TA., Shankar RA., Wightman PD., Reddy LG. The (NCBI) Human TLR8 "isoform 1" (1059 aa-NP_057694.2 ) used here, corresponds to UniProt TLR8 isoform 2. lgreddy@mmm.com 27-Mar-2018: 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 Wang J., Shao Y., Bennett TA., Shankar RA., Wightman PD., Reddy LG. The (NCBI) Human TLR8 "isoform 1" (1059 aa-NP_057694.2 ) used here, corresponds to UniProt TLR8 isoform 2. lgreddy@mmm.com Accepted 2018-MAR-15 AT 08:35 GMT AT 08:35 GMT by LIVIAP The Journal of biological chemistry 2006 Only protein-protein interactions imex curation imex curation The HA tag is not at the "N-terminUS" - it is "n-terminAL" according to text :- "The TLR8 expression vector with N-terminal HA tag was constructed by inserting the HA tag at the C-terminal end of the predicted signal peptide sequence (after 45 residues from the N-terminal end or TLR8 isoform 1)". I think they mean "OF TLR8 isoform 1" not "or isoform 1". I described the tag as "N-terminal" as they do in the text, as opposed to "N-terminus" which I would use if the tag was attached, as usual, at aas 1-~3 of a sequence. tlr7_human Toll-like receptor 7 TLR7 UNQ248/PRO285 protein protein human Homo sapiens Human MVFPMWTLKRQILILFNIILISKLLGARWFPKTLPCDVTLDVPKNHVIVDCTDKHLTEIPGGIPTNTTNLTLTINHIPDISPASFHRLDHLVEIDFRCNCVPIPLGSKNNMCIKRLQIKPRSFSGLTYLKSLYLDGNQLLEIPQGLPPSLQLLSLEANNIFSIRKENLTELANIEILYLGQNCYYRNPCYVSYSIEKDAFLNLTKLKVLSLKDNNVTAVPTVLPSTLTELYLYNNMIAKIQEDDFNNLNQLQILDLSGNCPRCYNAPFPCAPCKNNSPLQIPVNAFDALTELKVLRLHSNSLQHVPPRWFKNINKLQELDLSQNFLAKEIGDAKFLHFLPSLIQLDLSFNFELQVYRASMNLSQAFSSLKSLKILRIRGYVFKELKSFNLSPLHNLQNLEVLDLGTNFIKIANLSMFKQFKRLKVIDLSVNKISPSGDSSEVGFCSNARTSVESYEPQVLEQLHYFRYDKYARSCRFKNKEASFMSVNESCYKYGQTLDLSKNSIFFVKSSDFQHLSFLKCLNLSGNLISQTLNGSEFQPLAELRYLDFSNNRLDLLHSTAFEELHKLEVLDISSNSHYFQSEGITHMLNFTKNLKVLQKLMMNDNDISSSTSRTMESESLRTLEFRGNHLDVLWREGDNRYLQLFKNLLKLEELDISKNSLSFLPSGVFDGMPPNLKNLSLAKNGLKSFSWKKLQCLKNLETLDLSHNQLTTVPERLSNCSRSLKNLILKNNQIRSLTKYFLQDAFQLRYLDLSSNKIQMIQKTSFPENVLNNLKMLLLHHNRFLCTCDAVWFVWWVNHTEVTIPYLATDVTCVGPGAHKGQSVISLDLYTCELDLTNLILFSLSISVSLFLMVMMTASHLYFWDVWYIYHFCKAKIKGYQRLISPDCCYDAFIVYDTKDPAVTEWVLAELVAKLEDPREKHFNLCLEERDWLPGQPVLENLSQSIQLSKKTVFVMTDKYAKTENFKIAFYLSHQRLMDEKVDVIILIFLEKPFQKSKFLQLRKRLCGSSVLEWPTNPQAHPYFWQCLKNALATDNHVAYSQVFKETV 8C701E9E437F2721 q9nr97-2 Toll-like receptor 8 TLR8 UNQ249/PRO286 protein protein human Homo sapiens Human MKESSLQNSSCSLGKETKKENMFLQSSMLTCIFLLISGSCELCAEENFSRSYPCDEKKQNDSVIAECSNRRLQEVPQTVGKYVTELDLSDNFITHITNESFQGLQNLTKINLNHNPNVQHQNGNPGIQSNGLNITDGAFLNLKNLRELLLEDNQLPQIPSGLPESLTELSLIQNNIYNITKEGISRLINLKNLYLAWNCYFNKVCEKTNIEDGVFETLTNLELLSLSFNSLSHVPPKLPSSLRKLFLSNTQIKYISEEDFKGLINLTLLDLSGNCPRCFNAPFPCVPCDGGASINIDRFAFQNLTQLRYLNLSSTSLRKINAAWFKNMPHLKVLDLEFNYLVGEIASGAFLTMLPRLEILDLSFNYIKGSYPQHINISRNFSKLLSLRALHLRGYVFQELREDDFQPLMQLPNLSTINLGINFIKQIDFKLFQNFSNLEIIYLSENRISPLVKDTRQSYANSSSFQRHIRKRRSTDFEFDPHSNFYHFTRPLIKPQCAAYGKALDLSLNSIFFIGPNQFENLPDIACLNLSANSNAQVLSGTEFSAIPHVKYLDLTNNRLDFDNASALTELSDLEVLDLSYNSHYFRIAGVTHHLEFIQNFTNLKVLNLSHNNIYTLTDKYNLESKSLVELVFSGNRLDILWNDDDNRYISIFKGLKNLTRLDLSLNRLKHIPNEAFLNLPASLTELHINDNMLKFFNWTLLQQFPRLELLDLRGNKLLFLTDSLSDFTSSLRTLLLSHNRISHLPSGFLSEVSSLKHLDLSSNLLKTINKSALETKTTTKLSMLELHGNPFECTCDIGDFRRWMDEHLNVKIPRLVDVICASPGDQRGKSIVSLELTTCVSDVTAVILFFFTFFITTMVMLAALAHHLFYWDVWFIYNVCLAKVKGYRSLSTSQTFYDAYISYDTKDASVTDWVINELRYHLEESRDKNVLLCLEERDWDPGLAIIDNLMQSINQSKKTVFVLTKKYAKSWNFKTAFYLALQRLMDENMDVIIFILLEPVLQHSQYLRLRQRICKSSILQWPDNPKAEGLFWQTLRNVVLTENDSRYNNMYVDSIKQY 27803D560E09D7BF tlr9_human Toll-like receptor 9 TLR9 UNQ5798/PRO19605 protein protein human Homo sapiens Human MGFCRSALHPLSLLVQAIMLAMTLALGTLPAFLPCELQPHGLVNCNWLFLKSVPHFSMAAPRGNVTSLSLSSNRIHHLHDSDFAHLPSLRHLNLKWNCPPVGLSPMHFPCHMTIEPSTFLAVPTLEELNLSYNNIMTVPALPKSLISLSLSHTNILMLDSASLAGLHALRFLFMDGNCYYKNPCRQALEVAPGALLGLGNLTHLSLKYNNLTVVPRNLPSSLEYLLLSYNRIVKLAPEDLANLTALRVLDVGGNCRRCDHAPNPCMECPRHFPQLHPDTFSHLSRLEGLVLKDSSLSWLNASWFRGLGNLRVLDLSENFLYKCITKTKAFQGLTQLRKLNLSFNYQKRVSFAHLSLAPSFGSLVALKELDMHGIFFRSLDETTLRPLARLPMLQTLRLQMNFINQAQLGIFRAFPGLRYVDLSDNRISGASELTATMGEADGGEKVWLQPGDLAPAPVDTPSSEDFRPNCSTLNFTLDLSRNNLVTVQPEMFAQLSHLQCLRLSHNCISQAVNGSQFLPLTGLQVLDLSHNKLDLYHEHSFTELPRLEALDLSYNSQPFGMQGVGHNFSFVAHLRTLRHLSLAHNNIHSQVSQQLCSTSLRALDFSGNALGHMWAEGDLYLHFFQGLSGLIWLDLSQNRLHTLLPQTLRNLPKSLQVLRLRDNYLAFFKWWSLHFLPKLEVLDLAGNQLKALTNGSLPAGTRLRRLDVSCNSISFVAPGFFSKAKELRELNLSANALKTVDHSWFGPLASALQILDVSANPLHCACGAAFMDFLLEVQAAVPGLPSRVKCGSPGQLQGLSIFAQDLRLCLDEALSWDCFALSLLAVALGLGVPMLHHLCGWDLWYCFHLCLAWLPWRGRQSGRDEDALPYDAFVVFDKTQSAVADWVYNELRGQLEECRGRWALRLCLEERDWLPGKTLFENLWASVYGSRKTLFVLAHTDRVSGLLRASFLLAQQRLLEDRKDVVVLVILSPDGRRSRYVRLRQRLCRQSVLLWPHQPSGQRSFWAQLGMALTRDNHHFYNRNFCQGPTAE 71280AA9680EDCE2 tlr8-tlr7-1 2 4 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed n_terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag certain certain sequence position certain certain certain sequence position certain 3 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed c_terminus egfp tag enhanced green fluorescent protein tag enhanced green fluorescent protein EGFP enhanced green fluorescent protein EGFP egfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range physical association physical association Figure 3B tlr8-tlr7-2 1 4 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed n_terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag certain certain sequence position certain certain certain sequence position certain 5 unspecified role unspecified role prey prey physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association Figure 3C HA-TLR8 pull down using anti-HA antibody co-IPs TLR7-EGFP (Fig. 3B) and TLR9