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/ liu-2001-7 Direct inhibition of Bruton's tyrosine kinase by IBtk, a Btk-binding protein. in vitro In vitro protein kinase assay protein kinase assay predetermined predetermined participant predetermined Liu W., Quinto I., Chen X., Palmieri C., Rabin RL., Schwartz OM., Nelson DL., Scala G. Apoptosis - Interactions involving proteins with a function related to apoptosis scala@dbbm.unina.it A human B cell cDNA library was used. Nat. Immunol. (1529-2908) 2001 Only protein-protein interactions imex curation Cancer - Interactions investigated in the context of cancer liu-2001-1 Direct inhibition of Bruton's tyrosine kinase by IBtk, a Btk-binding protein. yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Liu W., Quinto I., Chen X., Palmieri C., Rabin RL., Schwartz OM., Nelson DL., Scala G. Apoptosis - Interactions involving proteins with a function related to apoptosis scala@dbbm.unina.it A human B cell cDNA library was used. Nat. Immunol. (1529-2908) 2001 Only protein-protein interactions imex curation Cancer - Interactions investigated in the context of cancer liu-2001-2 Direct inhibition of Bruton's tyrosine kinase by IBtk, a Btk-binding protein. in vitro In vitro pull down pull down weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Liu W., Quinto I., Chen X., Palmieri C., Rabin RL., Schwartz OM., Nelson DL., Scala G. Apoptosis - Interactions involving proteins with a function related to apoptosis scala@dbbm.unina.it A human B cell cDNA library was used. Nat. Immunol. (1529-2908) 2001 Only protein-protein interactions imex curation Cancer - Interactions investigated in the context of cancer liu-2001-3 Direct inhibition of Bruton's tyrosine kinase by IBtk, a Btk-binding protein. human-b-cell Homo sapiens B cell b-cell B-cell [B lymphocyte; B-lymphocyte; B cell; B-cells] pull down pull down western blot western blot Immuno blot Immuno blot Liu W., Quinto I., Chen X., Palmieri C., Rabin RL., Schwartz OM., Nelson DL., Scala G. Apoptosis - Interactions involving proteins with a function related to apoptosis scala@dbbm.unina.it A human B cell cDNA library was used. Nat. Immunol. (1529-2908) 2001 Only protein-protein interactions imex curation Cancer - Interactions investigated in the context of cancer liu-2001-4 Direct inhibition of Bruton's tyrosine kinase by IBtk, a Btk-binding protein. human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Liu W., Quinto I., Chen X., Palmieri C., Rabin RL., Schwartz OM., Nelson DL., Scala G. Apoptosis - Interactions involving proteins with a function related to apoptosis scala@dbbm.unina.it A human B cell cDNA library was used. Nat. Immunol. (1529-2908) 2001 Only protein-protein interactions imex curation Cancer - Interactions investigated in the context of cancer liu-2001-5 Direct inhibition of Bruton's tyrosine kinase by IBtk, a Btk-binding protein. human-b-cell Homo sapiens B cell b-cell B-cell [B lymphocyte; B-lymphocyte; B cell; B-cells] anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Liu W., Quinto I., Chen X., Palmieri C., Rabin RL., Schwartz OM., Nelson DL., Scala G. Apoptosis - Interactions involving proteins with a function related to apoptosis A web search revealed that DeFew cells are a human B lymphoma cell line. PMID 7787756 states that MC3 cells are a chronic myeloid leukemia cell line. This experiment covers the data regarding MC3 and DeFew cells in Figure 3 E. This data has been merged as both cell lines are B cell lines. scala@dbbm.unina.it A human B cell cDNA library was used. Nat. Immunol. (1529-2908) 2001 Only protein-protein interactions imex curation Cancer - Interactions investigated in the context of cancer liu-2001-6 Direct inhibition of Bruton's tyrosine kinase by IBtk, a Btk-binding protein. human-b-cell Homo sapiens B cell b-cell B-cell [B lymphocyte; B-lymphocyte; B cell; B-cells] confocal microscopy confocal microscopy immunostaining immunostaining Liu W., Quinto I., Chen X., Palmieri C., Rabin RL., Schwartz OM., Nelson DL., Scala G. Apoptosis - Interactions involving proteins with a function related to apoptosis scala@dbbm.unina.it A human B cell cDNA library was used. Nat. Immunol. (1529-2908) 2001 Only protein-protein interactions imex curation Cancer - Interactions investigated in the context of cancer btk_human Tyrosine-protein kinase BTK Bruton tyrosine kinase B-cell progenitor kinase BPK BTK AGMX1 ATK Agammaglobulinemia tyrosine kinase protein protein human Homo sapiens Human MAAVILESIFLKRSQQKKKTSPLNFKKRLFLLTVHKLSYYEYDFERGRRGSKKGSIDVEKITCVETVVPEKNPPPERQIPRRGEESSEMEQISIIERFPYPFQVVYDEGPLYVFSPTEELRKRWIHQLKNVIRYNSDLVQKYHPCFWIDGQYLCCSQTAKNAMGCQILENRNGSLKPGSSHRKTKKPLPPTPEEDQILKKPLPPEPAAAPVSTSELKKVVALYDYMPMNANDLQLRKGDEYFILEESNLPWWRARDKNGQEGYIPSNYVTEAEDSIEMYEWYSKHMTRSQAEQLLKQEGKEGGFIVRDSSKAGKYTVSVFAKSTGDPQGVIRHYVVCSTPQSQYYLAEKHLFSTIPELINYHQHNSAGLISRLKYPVSQQNKNAPSTAGLGYGSWEIDPKDLTFLKELGTGQFGVVKYGKWRGQYDVAIKMIKEGSMSEDEFIEEAKVMMNLSHEKLVQLYGVCTKQRPIFIITEYMANGCLLNYLREMRHRFQTQQLLEMCKDVCEAMEYLESKQFLHRDLAARNCLVNDQGVVKVSDFGLSRYVLDDEYTSSVGSKFPVRWSPPEVLMYSKFSSKSDIWAFGVLMWEIYSLGKMPYERFTNSETAEHIAQGLRLYRPHLASEKVYTIMYSCWHEKADERPTFKILLSNILDVMDEES ebi-1521221 Inhibitor of Bruton tyrosine kinase protein protein human Homo sapiens Human MLIDIISSKMIALTTKENNSGMNSMETVLFTPSKAPKPVNAWASSLHSVSSKSFRDFLLEEKKSVTSHSSGDHVKKVSFKGIENSQAPKIVRCSTHGTPGPEGNHISDLPLLDSPNPWLSSSVTAPSMVAPVTFASIVEEELQQEAALIRSREKPLALIQIEEHAIQDLLVFYEAFGNPEEFVIVERTPQGPLAVPMWNKHGC The sequence has been withdrawn from uniprot. sequence is significantly different from parent. ibtk_human Inhibitor of Bruton tyrosine kinase IBTK BTKI KIAA1417 protein protein human Homo sapiens Human MSSPMPDCTSKCRSLKHALDVLSVVTKGSENQIKAFLSSHCYNAATIKDVFGRNALHLVSSCGKKGVLDWLIQKGVDLLVKDKESGWTALHRSIFYGHIDCVWSLLKHGVSLYIQDKEGLSALDLVMKDRPTHVVFKNTDPTDVYTWGDNTNFTLGHGSQNSKHHPELVDLFSRSGIYIKQVVLCKFHSVFLSQKGQVYTCGHGPGGRLGHGDEQTCLVPRLVEGLNGHNCSQVAAAKDHTVVLTEDGCVYTFGLNIFHQLGIIPPPSSCNVPRQIQAKYLKGRTIIGVAAGRFHTVLWTREAVYTMGLNGGQLGCLLDPNGEKCVTAPRQVSALHHKDIALSLVAASDGATVCVTTRGDIYLLADYQCKKMASKQLNLKKVLVSGGHMEYKVDPEHLKENGGQKICILAMDGAGRVFCWRSVNSSLKQCRWAYPRQVFISDIALNRNEILFVTQDGEGFRGRWFEEKRKSSEKKEILSNLHNSSSDVSYVSDINSVYERIRLEKLTFAHRAVSVSTDPSGCNFAILQSDPKTSLYEIPAVSSSSFFEEFGKLLREADEMDSIHDVTFQVGNRLFPAHKYILAVHSDFFQKLFLSDGNTSEFTDIYQKDEDSAGCHLFVVEKVHPDMFEYLLQFIYTDTCDFLTHGFKPRIHLNKNPEEYQGTLNSHLNKVNFHEDDNQKSAFEVYKSNQAQTVSERQKSKPKSCKKGKNIREDDPVRMLQTVAKKFDFSNLSSRLDGVRFENEKINVIAKNTGNKLKLSQKKCSFLCDVTMKSVDGKEFPCHKCVLCARLEYFHSMLSSSWIEASSCAALEMPIHSDILKVILDYLYTDEAVVIKESQNVDFICSVLVVADQLLITRLKEICEVALTEKLTLKNAAMLLEFAAMYSAKQLKLSCLQFIGLNMAALLEARSLDVLSDGVLKDLSEFYRKMIPAMDRRVITPYQDGPDISYLEVEDGDIFLKEEINMEQNHSETMFKKAKTKAKKKPRKRSDSSGGYNLSDIIQSPSSTGLLKSGKTNSVESLPELLTSDSEGSYAGVGSPRDLQSPDFTTGFHSDKIEAKVKPYVNGTSPVYSREDLKPWEKSPILKISAPQPIPSNRIDTTSSASWVAGSFSPVSPPVVDLRTIMEIEESRQKCGATPKSHLGKTVSHGVKLSQKQRKMIALTTKENNSGMNSMETVLFTPSKAPKPVNAWASSLHSVSSKSFRDFLLEEKKSVTSHSSGDHVKKVSFKGIENSQAPKIVRCSTHGTPGPEGNHISDLPLLDSPNPWLSSSVTAPSMVAPVTFASIVEEELQQEAALIRSREKPLALIQIEEHAIQDLLVFYEAFGNPEEFVIVERTPQGPLAVPMWNKHGC btk-btk_substrat GST-BTK phosphorylation of BTK substrate peptide is inhibited by IBTK 729117 N/A 729124 enzyme target enzyme target substrate substrate unspecified role unspecified role region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) N/A 729124 enzyme enzyme unspecified role unspecified role region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation false false false 4 A IBTK - GST-IBTK inhibition of BTK kinase activity increases with increasing concentration of GST-IBTK. The BTK substrate peptide (BTK 217-229) is autophosphorylated on Tyr-223. btk-btk_substrat-1 BTK phosphorylation of BTK substrate peptide is inhibited by IBTK 729117 N/A 729124 enzyme enzyme unspecified role unspecified role human-b-cell Homo sapiens B cell b-cell B-cell [B lymphocyte; B-lymphocyte; B cell; B-cells] N/A 729124 enzyme target enzyme target substrate substrate unspecified role unspecified role region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) phosphorylation phosphorylation reaction phosphorylation false false false 4 C, E, F IBTK - GST-IBTK inhibition of BTK kinase activity increases with increasing concentration of GST-IBTK. Deletion analysis revealed that the C-terminal construct of IBTK retained inhibitory activty but the N-terminal construct did not. The BTK substrate peptide (BTK 217-229) is autophosphorylated on Tyr-223. IBTK causes nearly complete inhibition of BTK from unstimulated cells and only a substantial decrease in BCR-stimulated cells. btk-ibtk BTK and IBTK interact in a yeast two-hybrid assay 729118 N/A 729125 unspecified role unspecified role prey prey N/A 729124 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind range range range range false physical association physical association false false false The coordinates given for the BTK construct used, do not match with the length of the protein nor the coordinates of the specified domains as given by the UniProtKB entry. The range given is taken from the ranges of these domains in the UniProtKB entry. 1 btk-ibtk-1 GST-BTK pulls down IBTK in vitro 729119 N/A 729124 unspecified role unspecified role bait bait region PHBM sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis range range range range false region SH2 sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis range range range range false region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 729125 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false in vitro In vitro direct interaction direct interaction false false false The coordinates given for the BTK construct used, do not match with the coordinates of the specified domains as given by the UniProtKB entry. The ranges given are taken from the ranges of these domains in the UniProtKB entry. 3 A ibtk-btk GST-IBTK pulls down BTK from B cell lysate 729120 N/A 729124 unspecified role unspecified role prey prey human Homo sapiens Human N/A 729125 unspecified role unspecified role bait bait region gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false 3 C ibtk-btk-1 IBTK coimmunoprecipitates BTK in 293T lysates 729121 N/A 729124 unspecified role unspecified role prey prey human Homo sapiens Human N/A 729125 unspecified role unspecified role bait bait physical association physical association false false false 3 D Q9P2D0-4; Q06187 transfection IBTK peptide - An IBTK peptide displaced the immunoprecipitation activity of the anti-IBTK serum. ibtk-btk-2 IBTK coimmunoprecipitates BTK in B cell lysates 729122 N/A 729124 unspecified role unspecified role prey prey N/A 729126 unspecified role unspecified role bait bait physical association physical association false false false 3 E btk-ibtk-2 IBTK colocalizes BTK in MC3 lysates 729123 N/A 729126 unspecified role unspecified role neutral component neutral component N/A 729124 unspecified role unspecified role neutral component neutral component colocalization colocalization false false false 3 F, G In unstimulated cells, colocalization of IBtk and Btk took place in the cytoplasm. Upon BCR cross-linking, colocalization took place in the sub-membrane region.