bhf-ucl cory-1996-2 Evidence that the Wiskott-Aldrich syndrome protein may be involved in lymphoid cell signaling pathways. 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 J. Immunol. (0022-1767) 1996 imex curation Only protein-protein interactions cory-1996-1 Evidence that the Wiskott-Aldrich syndrome protein may be involved in lymphoid cell signaling pathways. 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 J. Immunol. (0022-1767) 1996 imex curation Only protein-protein interactions 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 wasp_human Wiskott-Aldrich syndrome protein WAS IMD2 protein protein human Homo sapiens Human MSGGPMGGRPGGRGAPAVQQNIPSTLLQDHENQRLFEMLGRKCLTLATAVVQLYLALPPGAEHWTKEHCGAVCFVKDNPQKSYFIRLYGLQAGRLLWEQELYSQLVYSTPTPFFHTFAGDDCQAGLNFADEDEAQAFRALVQEKIQKRNQRQSGDRRQLPPPPTPANEERRGGLPPLPLHPGGDQGGPPVGPLSLGLATVDIQNPDITSSRYRGLPAPGPSPADKKRSGKKKISKADIGAPSGFKHVSHVGWDPQNGFDVNNLDPDLRSLFSRAGISEAQLTDAETSKLIYDFIEDQGGLEAVRQEMRRQEPLPPPPPPSRGGNQLPRPPIVGGNKGRSGPLPPVPLGIAPPPPTPRGPPPPGRGGPPPPPPPATGRSGPLPPPPPGAGGPPMPPPPPPPPPPPSSGNGPAPPPLPPALVPAGGLAPGGGRGALLDQIRQGIQLNKTPGAPESSALQPPPQSSEGLVGALMHVMQKRSRAIHSSDEGEDQAGDEDEDDEWDD tec_human Tyrosine-protein kinase Tec PSCTK4 TEC protein protein human Homo sapiens Human MNFNTILEEILIKRSQQKKKTSPLNYKERLFVLTKSMLTYYEGRAEKKYRKGFIDVSKIKCVEIVKNDDGVIPCQNKYPFQVVHDANTLYIFAPSPQSRDLWVKKLKEEIKNNNNIMIKYHPKFWTDGSYQCCRQTEKLAPGCEKYNLFESSIRKALPPAPETKKRRPPPPIPLEEEDNSEEIVVAMYDFQAAEGHDLRLERGQEYLILEKNDVHWWRARDKYGNEGYIPSNYVTGKKSNNLDQYEWYCRNMNRSKAEQLLRSEDKEGGFMVRDSSQPGLYTVSLYTKFGGEGSSGFRHYHIKETTTSPKKYYLAEKHAFGSIPEIIEYHKHNAAGLVTRLRYPVSVKGKNAPTTAGFSYEKWEINPSELTFMRELGSGLFGVVRLGKWRAQYKVAIKAIREGAMCEEDFIEEAKVMMKLTHPKLVQLYGVCTQQKPIYIVTEFMERGCLLNFLRQRQGHFSRDVLLSMCQDVCEGMEYLERNSFIHRDLAARNCLVSEAGVVKVSDFGMARYVLDDQYTSSSGAKFPVKWCPPEVFNYSRFSSKSDVWSFGVLMWEVFTEGRMPFEKYTNYEVVTMVTRGHRLYQPKLASNYVYEVMLRCWQEKPEGRPSFEDLLRTIDELVECEETFGR 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 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 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 btk-was-1 1564275 n/a 1564277 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 sh3 domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1564278 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 false false false Fig 2 lane 2 btk-was-2 1564275 n/a 1564278 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 n/a 1564279 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 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 ecoli Escherichia coli (strain K12) physical association physical association false false false Fig 2 lane 6 tec-was 1564275 n/a 1564280 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 sh3 domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1564278 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 false false false Fig 2 lane 7 btk-was-3 1564275 n/a 1564281 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 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 ecoli Escherichia coli (strain K12) n/a 1564278 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 false false false Fig 2 lane 3 btk-was-4 1564275 n/a 1564282 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 sh3 domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1564278 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 false false false Fig 2 lane 5 btk-was-9 1564275 n/a WP2 1564278 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 false synthetic construct synthetic construct n/a 1564277 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 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 ecoli Escherichia coli (strain K12) n/a 1564278 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 1564312 1564310 physical association physical association false false false Fig 5 WP2 section btk-was-10 1564275 n/a 1564277 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 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 ecoli Escherichia coli (strain K12) n/a 1564278 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 n/a WP4 1564278 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 false synthetic construct synthetic construct 1564321 1564317 physical association physical association false false false Fig 5 WP4 section btk-was 1564276 n/a 1564277 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 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 ecoli Escherichia coli (strain K12) n/a 1564278 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 false false false Table 2 and Fig 1