bhf-ucl Evidence that the Wiskott-Aldrich syndrome protein may be involved in lymphoid cell signaling pathways. Evidence that the Wiskott-Aldrich syndrome protein may be involved in lymphoid cell signaling pathways. J. Immunol. (0022-1767) 1996 imex curation Cory G.O., MacCarthy-Morrogh L., Banin S., Gout I., Brickell P.M., Levinsky R.J., Kinnon C., Lovering R.C. Cardiac - Interactions involving cardiac related proteins All constructs were human (confirmed by author) r.lovering@ucl.ac.uk. Ref 20: PMID:7629518 also confirms human BTK GST SH3 domain fusion construct: nucleotides 787-951, with five additional aa at COOH-terminal, by PCR using primers SH3F (5'-CTGAGGATCCGTTGCCTTTATGATTAC) and SH3R (5'-TATCAGAATTCTGCTTCAGTGACATAGTTACTAG). The alignment of the primers with BTK ENST00000308731 and BTK Q06187 suggests the amplified region is 819-1023 so that SH3 domain is included in the fusion protein, covering 219-277 (the SH3 domain is 214-274) so very slight difference. Ref27 PMID:8402898 confirms human constructs for GRB2 and PLCgamma and defines the regions used. Gout paper refers to Kohda et al PMID:7681365 for PLCgamma, this paper is associated with the PLCG1 in UniProt therefore this gene annotated. Other SH3 construct regions calculated based on PCR primers presented in this paper. r.lovering@ucl.ac.uk 12-Nov-2014: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro pull down pull down edman degradation edman degradation Cory G.O., MacCarthy-Morrogh L., Banin S., Gout I., Brickell P.M., Levinsky R.J., Kinnon C., Lovering R.C. Cardiac - Interactions involving cardiac related proteins All constructs were human (confirmed by author) r.lovering@ucl.ac.uk. Ref 20: PMID:7629518 also confirms human BTK GST SH3 domain fusion construct: nucleotides 787-951, with five additional aa at COOH-terminal, by PCR using primers SH3F (5'-CTGAGGATCCGTTGCCTTTATGATTAC) and SH3R (5'-TATCAGAATTCTGCTTCAGTGACATAGTTACTAG). The alignment of the primers with BTK ENST00000308731 and BTK Q06187 suggests the amplified region is 819-1023 so that SH3 domain is included in the fusion protein, covering 219-277 (the SH3 domain is 214-274) so very slight difference. Ref27 PMID:8402898 confirms human constructs for GRB2 and PLCgamma and defines the regions used. Gout paper refers to Kohda et al PMID:7681365 for PLCgamma, this paper is associated with the PLCG1 in UniProt therefore this gene annotated. Other SH3 construct regions calculated based on PCR primers presented in this paper. r.lovering@ucl.ac.uk Accepted 2014-OCT-30 by ORCHARD J. Immunol. (0022-1767) 1996 imex curation Evidence that the Wiskott-Aldrich syndrome protein may be involved in lymphoid cell signaling pathways. Evidence that the Wiskott-Aldrich syndrome protein may be involved in lymphoid cell signaling pathways. J. Immunol. (0022-1767) 1996 imex curation Cory G.O., MacCarthy-Morrogh L., Banin S., Gout I., Brickell P.M., Levinsky R.J., Kinnon C., Lovering R.C. Cardiac - Interactions involving cardiac related proteins All constructs were human (confirmed by author) r.lovering@ucl.ac.uk. Ref 20: PMID:7629518 also confirms human BTK GST SH3 domain fusion construct: nucleotides 787-951, with five additional aa at COOH-terminal, by PCR using primers SH3F (5'-CTGAGGATCCGTTGCCTTTATGATTAC) and SH3R (5'-TATCAGAATTCTGCTTCAGTGACATAGTTACTAG). The alignment of the primers with BTK ENST00000308731 and BTK Q06187 suggests the amplified region is 819-1023 so that SH3 domain is included in the fusion protein, covering 219-277 (the SH3 domain is 214-274) so very slight difference. Ref27 PMID:8402898 confirms human constructs for GRB2 and PLCgamma and defines the regions used. Gout paper refers to Kohda et al PMID:7681365 for PLCgamma, this paper is associated with the PLCG1 in UniProt therefore this gene annotated. Other SH3 construct regions calculated based on PCR primers presented in this paper. r.lovering@ucl.ac.uk 12-Nov-2014: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro pull down pull down polyclonal western polyclonal antibody western blot polyclonal western Cory G.O., MacCarthy-Morrogh L., Banin S., Gout I., Brickell P.M., Levinsky R.J., Kinnon C., Lovering R.C. Cardiac - Interactions involving cardiac related proteins All constructs were human (confirmed by author) r.lovering@ucl.ac.uk. Ref 20: PMID:7629518 also confirms human BTK GST SH3 domain fusion construct: nucleotides 787-951, with five additional aa at COOH-terminal, by PCR using primers SH3F (5'-CTGAGGATCCGTTGCCTTTATGATTAC) and SH3R (5'-TATCAGAATTCTGCTTCAGTGACATAGTTACTAG). The alignment of the primers with BTK ENST00000308731 and BTK Q06187 suggests the amplified region is 819-1023 so that SH3 domain is included in the fusion protein, covering 219-277 (the SH3 domain is 214-274) so very slight difference. Ref27 PMID:8402898 confirms human constructs for GRB2 and PLCgamma and defines the regions used. Gout paper refers to Kohda et al PMID:7681365 for PLCgamma, this paper is associated with the PLCG1 in UniProt therefore this gene annotated. Other SH3 construct regions calculated based on PCR primers presented in this paper. r.lovering@ucl.ac.uk Accepted 2014-OCT-30 by ORCHARD J. Immunol. (0022-1767) 1996 imex curation tec_human Tyrosine-protein kinase Tec PSCTK4 TEC protein protein human Homo sapiens Human MNFNTILEEILIKRSQQKKKTSPLNYKERLFVLTKSMLTYYEGRAEKKYRKGFIDVSKIKCVEIVKNDDGVIPCQNKYPFQVVHDANTLYIFAPSPQSRDLWVKKLKEEIKNNNNIMIKYHPKFWTDGSYQCCRQTEKLAPGCEKYNLFESSIRKALPPAPETKKRRPPPPIPLEEEDNSEEIVVAMYDFQAAEGHDLRLERGQEYLILEKNDVHWWRARDKYGNEGYIPSNYVTGKKSNNLDQYEWYCRNMNRSKAEQLLRSEDKEGGFMVRDSSQPGLYTVSLYTKFGGEGSSGFRHYHIKETTTSPKKYYLAEKHAFGSIPEIIEYHKHNAAGLVTRLRYPVSVKGKNAPTTAGFSYEKWEINPSELTFMRELGSGLFGVVRLGKWRAQYKVAIKAIREGAMCEEDFIEEAKVMMKLTHPKLVQLYGVCTQQKPIYIVTEFMERGCLLNFLRQRQGHFSRDVLLSMCQDVCEGMEYLERNSFIHRDLAARNCLVSEAGVVKVSDFGMARYVLDDQYTSSSGAKFPVKWCPPEVFNYSRFSSKSDVWSFGVLMWEVFTEGRMPFEKYTNYEVVTMVTRGHRLYQPKLASNYVYEVMLRCWQEKPEGRPSFEDLLRTIDELVECEETFGR F55DECBF9916E1F9 itk_human Tyrosine-protein kinase ITK/TSK ITK EMT LYK T-cell-specific kinase Tyrosine-protein kinase Lyk Kinase EMT Interleukin-2-inducible T-cell kinase protein protein human Homo sapiens Human MNNFILLEEQLIKKSQQKRRTSPSNFKVRFFVLTKASLAYFEDRHGKKRTLKGSIELSRIKCVEIVKSDISIPCHYKYPFQVVHDNYLLYVFAPDRESRQRWVLALKEETRNNNSLVPKYHPNFWMDGKWRCCSQLEKLATGCAQYDPTKNASKKPLPPTPEDNRRPLWEPEETVVIALYDYQTNDPQELALRRNEEYCLLDSSEIHWWRVQDRNGHEGYVPSSYLVEKSPNNLETYEWYNKSISRDKAEKLLLDTGKEGAFMVRDSRTAGTYTVSVFTKAVVSENNPCIKHYHIKETNDNPKRYYVAEKYVFDSIPLLINYHQHNGGGLVTRLRYPVCFGRQKAPVTAGLRYGKWVIDPSELTFVQEIGSGQFGLVHLGYWLNKDKVAIKTIREGAMSEEDFIEEAEVMMKLSHPKLVQLYGVCLEQAPICLVFEFMEHGCLSDYLRTQRGLFAAETLLGMCLDVCEGMAYLEEACVIHRDLAARNCLVGENQVIKVSDFGMTRFVLDDQYTSSTGTKFPVKWASPEVFSFSRYSSKSDVWSFGVLMWEVFSEGKIPYENRSNSEVVEDISTGFRLYKPRLASTHVYQIMNHCWKERPEDRPAFSRLLRQLAEIAESGL DIP protein Q08881 original sequence version: 157 DIP protein Q08881 original sequence version: 93 DAE396BD2309319D grb2_human Growth factor receptor-bound protein 2 GRB2 ASH Adapter protein GRB2 SH2/SH3 adapter GRB2 Protein Ash protein protein human Homo sapiens Human MEAIAKYDFKATADDELSFKRGDILKVLNEECDQNWYKAELNGKDGFIPKNYIEMKPHPWFFGKIPRAKAEEMLSKQRHDGAFLIRESESAPGDFSLSVKFGNDVQHFKVLRDGAGKYFLWVVKFNSLNELVDYHRSTSVSRNQQIFLRDIEQVPQQPTYVQALFDFDPQEDGELGFRRGDFIHVMDNSDPNWWKGACHGQTGMFPRNYVTPVNRNV 83A4B0BA1B248DC4 wasp_human Wiskott-Aldrich syndrome protein WAS IMD2 protein protein human Homo sapiens Human MSGGPMGGRPGGRGAPAVQQNIPSTLLQDHENQRLFEMLGRKCLTLATAVVQLYLALPPGAEHWTKEHCGAVCFVKDNPQKSYFIRLYGLQAGRLLWEQELYSQLVYSTPTPFFHTFAGDDCQAGLNFADEDEAQAFRALVQEKIQKRNQRQSGDRRQLPPPPTPANEERRGGLPPLPLHPGGDQGGPPVGPLSLGLATVDIQNPDITSSRYRGLPAPGPSPADKKRSGKKKISKADIGAPSGFKHVSHVGWDPQNGFDVNNLDPDLRSLFSRAGISEAQLTDAETSKLIYDFIEDQGGLEAVRQEMRRQEPLPPPPPPSRGGNQLPRPPIVGGNKGRSGPLPPVPLGIAPPPPTPRGPPPPGRGGPPPPPPPATGRSGPLPPPPPGAGGPPMPPPPPPPPPPPSSGNGPAPPPLPPALVPAGGLAPGGGRGALLDQIRQGIQLNKTPGAPESSALQPPPQSSEGLVGALMHVMQKRSRAIHSSDEGEDQAGDEDEDDEWDD DIP protein P42768 original sequence version: 183 DIP protein P42768 original sequence version: 168 DIP protein P42768 original sequence version: 132 DIP protein P42768 original sequence version: 155 7228428672B7CB78 plcg1_human 1-phosphatidylinositol 4,5-bisphosphate phosphodiesterase gamma-1 PLC-148 PLC1 Phospholipase C-II PLCG1 Phosphoinositide phospholipase C-gamma-1 Phospholipase C-gamma-1 protein protein human Homo sapiens Human MAGAASPCANGCGPGAPSDAEVLHLCRSLEVGTVMTLFYSKKSQRPERKTFQVKLETRQITWSRGADKIEGAIDIREIKEIRPGKTSRDFDRYQEDPAFRPDQSHCFVILYGMEFRLKTLSLQATSEDEVNMWIKGLTWLMEDTLQAPTPLQIERWLRKQFYSVDRNREDRISAKDLKNMLSQVNYRVPNMRFLRERLTDLEQRSGDITYGQFAQLYRSLMYSAQKTMDLPFLEASTLRAGERPELCRVSLPEFQQFLLDYQGELWAVDRLQVQEFMLSFLRDPLREIEEPYFFLDEFVTFLFSKENSVWNSQLDAVCPDTMNNPLSHYWISSSHNTYLTGDQFSSESSLEAYARCLRMGCRCIELDCWDGPDGMPVIYHGHTLTTKIKFSDVLHTIKEHAFVASEYPVILSIEDHCSIAQQRNMAQYFKKVLGDTLLTKPVEISADGLPSPNQLKRKILIKHKKLAEGSAYEEVPTSMMYSENDISNSIKNGILYLEDPVNHEWYPHYFVLTSSKIYYSEETSSDQGNEDEEEPKEVSSSTELHSNEKWFHGKLGAGRDGRHIAERLLTEYCIETGAPDGSFLVRESETFVGDYTLSFWRNGKVQHCRIHSRQDAGTPKFFLTDNLVFDSLYDLITHYQQVPLRCNEFEMRLSEPVPQTNAHESKEWYHASLTRAQAEHMLMRVPRDGAFLVRKRNEPNSYAISFRAEGKIKHCRVQQEGQTVMLGNSEFDSLVDLISYYEKHPLYRKMKLRYPINEEALEKIGTAEPDYGALYEGRNPGFYVEANPMPTFKCAVKALFDYKAQREDELTFIKSAIIQNVEKQEGGWWRGDYGGKKQLWFPSNYVEEMVNPVALEPEREHLDENSPLGDLLRGVLDVPACQIAIRPEGKNNRLFVFSISMASVAHWSLDVAADSQEELQDWVKKIREVAQTADARLTEGKIMERRKKIALELSELVVYCRPVPFDEEKIGTERACYRDMSSFPETKAEKYVNKAKGKKFLQYNRLQLSRIYPKGQRLDSSNYDPLPMWICGSQLVALNFQTPDKPMQMNQALFMTGRHCGYVLQPSTMRDEAFDPFDKSSLRGLEPCAISIEVLGARHLPKNGRGIVCPFVEIEVAGAEYDSTKQKTEFVVDNGLNPVWPAKPFHFQISNPEFAFLRFVVYEEDMFSDQNFLAQATFPVKGLKTGYRAVPLKNNYSEDLELASLLIKIDIFPAKENGDLSPFSGTSLRERGSDASGQLFHGRAREGSFESRYQQPFEDFRISQEHLADHFDSRERRAPRRTRVNGDNRL DIP protein P19174 original sequence version: 144 DIP protein P19174 original sequence version: 152 DIP protein P19174 original sequence version: 181 DIP protein P19174 original sequence version: 169 AE05ABE2A18EDDAC 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 DIP protein Q06187 original sequence version: 118 DIP protein Q06187 original sequence version: 143 DF06B5D1FEC257CC btk-was 1 8 unspecified role unspecified role bait bait sh3 domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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 ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role prey prey human-daudi Homo sapiens Burkitt's B-cell lymphona cells daudi Homo sapies DAUDI Burkitt's B-cell lymphoma physical association physical association Table 2 and Fig 1 btk-was-1 2 8 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 sh3 domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role prey prey human-daudi Homo sapiens Burkitt's B-cell lymphona cells daudi Homo sapies DAUDI Burkitt's B-cell lymphoma physical association physical association Fig 2 lane 2 btk-was-2 2 6 unspecified role unspecified role prey prey human-daudi Homo sapiens Burkitt's B-cell lymphona cells daudi Homo sapies DAUDI Burkitt's B-cell lymphoma 3 unspecified role unspecified role bait bait sh3 domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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 ecoli Escherichia coli (strain K12) physical association physical association Fig 2 lane 6 tec-was 2 4 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 sh3 domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role prey prey human-daudi Homo sapiens Burkitt's B-cell lymphona cells daudi Homo sapies DAUDI Burkitt's B-cell lymphoma physical association physical association Fig 2 lane 7 btk-was-3 2 5 unspecified role unspecified role bait bait sh3 domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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 ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role prey prey human-daudi Homo sapiens Burkitt's B-cell lymphona cells daudi Homo sapies DAUDI Burkitt's B-cell lymphoma physical association physical association Fig 2 lane 3 btk-was-4 2 7 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 sh3 domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role prey prey human-daudi Homo sapiens Burkitt's B-cell lymphona cells daudi Homo sapies DAUDI Burkitt's B-cell lymphoma physical association physical association Fig 2 lane 5 btk-was-9 2 WP2 6 competitor competitor competitor prey prey purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain synthetic construct synthetic construct aa of peptides in table 1 match with uniprot numbering 8 unspecified role unspecified role bait bait sh3 domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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 ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role prey prey human-daudi Homo sapiens Burkitt's B-cell lymphona cells daudi Homo sapies DAUDI Burkitt's B-cell lymphoma aa of peptides in table 1 match with uniprot numbering 41 43 physical association physical association Fig 5 WP2 section btk-was-10 2 8 unspecified role unspecified role bait bait sh3 domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 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 ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role prey prey human-daudi Homo sapiens Burkitt's B-cell lymphona cells daudi Homo sapies DAUDI Burkitt's B-cell lymphoma aa of peptides in table 1 match with uniprot numbering WP4 6 competitor competitor competitor prey prey purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain synthetic construct synthetic construct aa of peptides in table 1 match with uniprot numbering 52 48 physical association physical association Fig 5 WP4 section