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/ Modulation of TLR4 signaling by a novel adaptor protein signal-transducing adaptor protein-2 in macrophages. Modulation of TLR4 signaling by a novel adaptor protein signal-transducing adaptor protein-2 in macrophages. J. Immunol. (0022-1767) 2006 imex curation Sekine Y., Yumioka T., Yamamoto T., Muromoto R., Imoto S., Sugiyma K., Oritani K., Shimoda K., Minoguchi M., Akira S., Yoshimura A., Matsuda T. The authors confirmed by email that the constructs for IKappaB and IKK-alpha were of human origin. tmatsuda@pharm.hokudai.ac.jp 22-Jan-2008: Contacted by JYOTI. DAVET Only protein-protein interactions human-thp-1 Homo sapiens acute monocytic leukemia cells thp-1 Human acute monocytic leukemia cell line anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Sekine Y., Yumioka T., Yamamoto T., Muromoto R., Imoto S., Sugiyma K., Oritani K., Shimoda K., Minoguchi M., Akira S., Yoshimura A., Matsuda T. The THP-1 cells were treated with PMA for 3 days to induce differentiation. The authors confirmed by email that the constructs for IKappaB and IKK-alpha were of human origin. tmatsuda@pharm.hokudai.ac.jp Accepted 2007-DEC-23 by DAVET. 22-Jan-2008: Contacted by JYOTI. J. Immunol. (0022-1767) 2006 DAVET Only protein-protein interactions imex curation Modulation of TLR4 signaling by a novel adaptor protein signal-transducing adaptor protein-2 in macrophages. Modulation of TLR4 signaling by a novel adaptor protein signal-transducing adaptor protein-2 in macrophages. J. Immunol. (0022-1767) 2006 imex curation Sekine Y., Yumioka T., Yamamoto T., Muromoto R., Imoto S., Sugiyma K., Oritani K., Shimoda K., Minoguchi M., Akira S., Yoshimura A., Matsuda T. The authors confirmed by email that the constructs for IKappaB and IKK-alpha were of human origin. tmatsuda@pharm.hokudai.ac.jp 22-Jan-2008: Contacted by JYOTI. DAVET Only protein-protein interactions in vitro In vitro protein kinase assay protein kinase assay weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Sekine Y., Yumioka T., Yamamoto T., Muromoto R., Imoto S., Sugiyma K., Oritani K., Shimoda K., Minoguchi M., Akira S., Yoshimura A., Matsuda T. 293T cells were transfected with His-tagged IKK-beta and/or the increasing amounts of Myc-tagged STAP-2 as indicated. Forty-eight hours after transfection, the cells were lysed, and immunoprecipitated with anti-His Ab. The kinase activity of IKK-beta in the immunoprecipitates was measured by in vitro kinase assay in the presence of GST-IkappaB. The authors confirmed by email that the constructs for IKappaB and IKK-alpha were of human origin. tmatsuda@pharm.hokudai.ac.jp Accepted 2007-DEC-23 by DAVET. 22-Jan-2008: Contacted by JYOTI. J. Immunol. (0022-1767) 2006 DAVET Only protein-protein interactions imex curation Modulation of TLR4 signaling by a novel adaptor protein signal-transducing adaptor protein-2 in macrophages. Modulation of TLR4 signaling by a novel adaptor protein signal-transducing adaptor protein-2 in macrophages. J. Immunol. (0022-1767) 2006 imex curation Sekine Y., Yumioka T., Yamamoto T., Muromoto R., Imoto S., Sugiyma K., Oritani K., Shimoda K., Minoguchi M., Akira S., Yoshimura A., Matsuda T. The authors confirmed by email that the constructs for IKappaB and IKK-alpha were of human origin. tmatsuda@pharm.hokudai.ac.jp 22-Jan-2008: Contacted by JYOTI. DAVET Only protein-protein interactions 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 tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Sekine Y., Yumioka T., Yamamoto T., Muromoto R., Imoto S., Sugiyma K., Oritani K., Shimoda K., Minoguchi M., Akira S., Yoshimura A., Matsuda T. The authors confirmed by email that the constructs for IKappaB and IKK-alpha were of human origin. tmatsuda@pharm.hokudai.ac.jp Accepted 2007-DEC-23 by DAVET. J. Immunol. (0022-1767) 2006 DAVET Only protein-protein interactions imex curation Modulation of TLR4 signaling by a novel adaptor protein signal-transducing adaptor protein-2 in macrophages. Modulation of TLR4 signaling by a novel adaptor protein signal-transducing adaptor protein-2 in macrophages. J. Immunol. (0022-1767) 2006 imex curation Sekine Y., Yumioka T., Yamamoto T., Muromoto R., Imoto S., Sugiyma K., Oritani K., Shimoda K., Minoguchi M., Akira S., Yoshimura A., Matsuda T. The authors confirmed by email that the constructs for IKappaB and IKK-alpha were of human origin. tmatsuda@pharm.hokudai.ac.jp 22-Jan-2008: Contacted by JYOTI. DAVET Only protein-protein interactions human-jurkat Homo sapiens jurkat Human T cell leukemia anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Sekine Y., Yumioka T., Yamamoto T., Muromoto R., Imoto S., Sugiyma K., Oritani K., Shimoda K., Minoguchi M., Akira S., Yoshimura A., Matsuda T. The Jurkat cells were stably transfected with Myc-STAP-2. The authors confirmed by email that the constructs for IKappaB and IKK-alpha were of human origin. tmatsuda@pharm.hokudai.ac.jp Accepted 2007-DEC-23 by DAVET. 22-Jan-2008: Contacted by JYOTI. J. Immunol. (0022-1767) 2006 DAVET Only protein-protein interactions imex curation Modulation of TLR4 signaling by a novel adaptor protein signal-transducing adaptor protein-2 in macrophages. Modulation of TLR4 signaling by a novel adaptor protein signal-transducing adaptor protein-2 in macrophages. J. Immunol. (0022-1767) 2006 imex curation Sekine Y., Yumioka T., Yamamoto T., Muromoto R., Imoto S., Sugiyma K., Oritani K., Shimoda K., Minoguchi M., Akira S., Yoshimura A., Matsuda T. The authors confirmed by email that the constructs for IKappaB and IKK-alpha were of human origin. tmatsuda@pharm.hokudai.ac.jp 22-Jan-2008: Contacted by JYOTI. DAVET Only protein-protein interactions 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. pull down pull down anti tag western anti tag western blot anti tag western Sekine Y., Yumioka T., Yamamoto T., Muromoto R., Imoto S., Sugiyma K., Oritani K., Shimoda K., Minoguchi M., Akira S., Yoshimura A., Matsuda T. The authors confirmed by email that the constructs for IKappaB and IKK-alpha were of human origin. tmatsuda@pharm.hokudai.ac.jp Accepted 2007-DEC-23 by DAVET. 22-Jan-2008: Contacted by JYOTI. J. Immunol. (0022-1767) 2006 DAVET Only protein-protein interactions imex curation myd88_human Myeloid differentiation primary response protein MyD88 MYD88 protein protein human Homo sapiens Human MAAGGPGAGSAAPVSSTSSLPLAALNMRVRRRLSLFLNVRTQVAADWTALAEEMDFEYLEIRQLETQADPTGRLLDAWQGRPGASVGRLLELLTKLGRDDVLLELGPSIEEDCQKYILKQQQEEAEKPLQVAAVDSSVPRTAELAGITTLDDPLGHMPERFDAFICYCPSDIQFVQEMIRQLEQTNYRLKLCVSDRDVLPGTCVWSIASELIEKRCRRMVVVVSDDYLQSKECDFQTKFALSLSPGAHQKRLIPIKYKAMKKEFPSILRFITVCDYTNPCTKSWFWTRLAKALSLP CEAE3F6B99524333 ikka_human Inhibitor of nuclear factor kappa-B kinase subunit alpha CHUK IKKA TCF16 I-kappa-B kinase 1 Conserved helix-loop-helix ubiquitous kinase Nuclear factor NF-kappa-B inhibitor kinase alpha Transcription factor 16 protein protein human Homo sapiens Human MERPPGLRPGAGGPWEMRERLGTGGFGNVCLYQHRELDLKIAIKSCRLELSTKNRERWCHEIQIMKKLNHANVVKACDVPEELNILIHDVPLLAMEYCSGGDLRKLLNKPENCCGLKESQILSLLSDIGSGIRYLHENKIIHRDLKPENIVLQDVGGKIIHKIIDLGYAKDVDQGSLCTSFVGTLQYLAPELFENKPYTATVDYWSFGTMVFECIAGYRPFLHHLQPFTWHEKIKKKDPKCIFACEEMSGEVRFSSHLPQPNSLCSLVVEPMENWLQLMLNWDPQQRGGPVDLTLKQPRCFVLMDHILNLKIVHILNMTSAKIISFLLPPDESLHSLQSRIERETGINTGSQELLSETGISLDPRKPASQCVLDGVRGCDSYMVYLFDKSKTVYEGPFASRSLSDCVNYIVQDSKIQLPIIQLRKVWAEAVHYVSGLKEDYSRLFQGQRAAMLSLLRYNANLTKMKNTLISASQQLKAKLEFFHKSIQLDLERYSEQMTYGISSEKMLKAWKEMEEKAIHYAEVGVIGYLEDQIMSLHAEIMELQKSPYGRRQGDLMESLEQRAIDLYKQLKHRPSDHSYSDSTEMVKIIVHTVQSQDRVLKELFGHLSKLLGCKQKIIDLLPKVEVALSNIKEADNTVMFMQGKRQKEIWHLLKIACTQSSARSLVGSSLEGAVTPQTSAWLPPTSAEHDHSLSCVVTPQDGETSAQMIEENLNCLGHLSTIIHEANEEQGNSMMNLDWSWLTE DIP protein O15111 original sequence version: 91 DIP protein O15111 original sequence version: 123 DIP protein O15111 original sequence version: 133 DIP protein O15111 original sequence version: 118 DIP protein O15111 original sequence version: 125 DIP protein O15111 original sequence version: 96 4EA55C6FFC66FA16 ikkb_human Inhibitor of nuclear factor kappa-B kinase subunit beta IKBKB IKKB I-kappa-B kinase 2 Nuclear factor NF-kappa-B inhibitor kinase beta Serine/threonine protein kinase IKBKB protein protein human Homo sapiens Human MSWSPSLTTQTCGAWEMKERLGTGGFGNVIRWHNQETGEQIAIKQCRQELSPRNRERWCLEIQIMRRLTHPNVVAARDVPEGMQNLAPNDLPLLAMEYCQGGDLRKYLNQFENCCGLREGAILTLLSDIASALRYLHENRIIHRDLKPENIVLQQGEQRLIHKIIDLGYAKELDQGSLCTSFVGTLQYLAPELLEQQKYTVTVDYWSFGTLAFECITGFRPFLPNWQPVQWHSKVRQKSEVDIVVSEDLNGTVKFSSSLPYPNNLNSVLAERLEKWLQLMLMWHPRQRGTDPTYGPNGCFKALDDILNLKLVHILNMVTGTIHTYPVTEDESLQSLKARIQQDTGIPEEDQELLQEAGLALIPDKPATQCISDGKLNEGHTLDMDLVFLFDNSKITYETQISPRPQPESVSCILQEPKRNLAFFQLRKVWGQVWHSIQTLKEDCNRLQQGQRAAMMNLLRNNSCLSKMKNSMASMSQQLKAKLDFFKTSIQIDLEKYSEQTEFGITSDKLLLAWREMEQAVELCGRENEVKLLVERMMALQTDIVDLQRSPMGRKQGGTLDDLEEQARELYRRLREKPRDQRTEGDSQEMVRLLLQAIQSFEKKVRVIYTQLSKTVVCKQKALELLPKVEEVVSLMNEDEKTVVRLQEKRQKELWNLLKIACSKVRGPVSGSPDSMNASRLSQPGQLMSQPSTASNSLPEPAKKSEELVAEAHNLCTLLENAIQDTVREQDQSFTALDWSWLQTEEEEHSCLEQAS DIP protein O14920 original sequence version: 89 DIP protein O14920 original sequence version: 100 DIP protein O14920 original sequence version: 121 DIP protein O14920 original sequence version: 126 DIP protein O14920 original sequence version: 136 DIP protein O14920 original sequence version: 128 DIP protein O14920-1 original sequence version: 151 DIP protein O14920 original sequence version: 142 DIP protein O14920 original sequence version: 159 DIP protein O14920 original sequence version: 93 DIP protein O14920 original sequence version: 124 DIP protein O14920-1 original sequence version: 178 DIP protein O14920 original sequence version: 85 F9CADF671AE9E14E stap2_human Signal-transducing adaptor protein 2 Breast tumor kinase substrate STAP2 BKS protein protein human Homo sapiens Human MASALRPPRVPKPKGVLPSHYYESFLEKKGPCDRDYKKFWAGLQGLTIYFYNSNRDFQHVEKLNLGAFEKLTDEIPWGSSRDPGTHFSLILRDQEIKFKVETLECREMWKGFILTVVELRVPTDLTLLPGHLYMMSEVLAKEEARRALETPSCFLKVSRLEAQLLLERYPECGNLLLRPSGDGADGVSVTTRQMHNGTHVVRHYKVKREGPKYVIDVEQPFSCTSLDAVVNYFVSHTKKALVPFLLDEDYEKVLGYVEADKENGENVWVAPSAPGPGPAPCTGGPKPLSPASSQDKLPPLPPLPNQEENYVTPIGDGPAVDYENQDVASSSWPVILKPKKLPKPPAKLPKPPVGPKPEPKVFNGGLGRKLPVSSAQPLFPTAGLADMTAELQKKLEKRRALEH A5E809B3F233EFD0 ikba_human NF-kappa-B inhibitor alpha NFKBIA IKBA MAD3 NFKBI I-kappa-B-alpha Major histocompatibility complex enhancer-binding protein MAD3 protein protein human Homo sapiens Human MFQAAERPQEWAMEGPRDGLKKERLLDDRHDSGLDSMKDEEYEQMVKELQEIRLEPQEVPRGSEPWKQQLTEDGDSFLHLAIIHEEKALTMEVIRQVKGDLAFLNFQNNLQQTPLHLAVITNQPEIAEALLGAGCDPELRDFRGNTPLHLACEQGCLASVGVLTQSCTTPHLHSILKATNYNGHTCLHLASIHGYLGIVELLVSLGADVNAQEPCNGRTALHLAVDLQNPDLVSLLLKCGADVNRVTYQGYSPYQLTWGRPSTRIQQQLGQLTLENLQMLPESEDEESYDTESEFTEFTEDELPYDDCVFGGQRLTL DIP protein P25963 original sequence version: 160 DIP protein P25963 original sequence version: 157 DIP protein P25963 original sequence version: 145 DIP protein P25963 original sequence version: 170 DIP protein P25963 original sequence version: 122 DIP protein P25963 original sequence version: 128 DIP protein P25963 original sequence version: 78 DIP protein P25963 original sequence version: 121 DIP protein P25963 original sequence version: 124 088B313226786395 stap2-myd88 3 6 unspecified role unspecified role prey prey region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 9 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association 3 B When expression vectors for FLAG-tagged MyD88 or His-tagged IKK- together with Myc-tagged STAP-2 were transfected into 293T cells and the cells were lysed, immunoprecipitated with an anti-Myc Ab, and immunoblotted with an anti-FLAG or anti-His Ab, the immunoprecipitates contained MyD88 and IKK- (Fig. 3, B and C), indicating that STAP-2 associated with MyD88 and IKK- in 293T cells. Q9UGK3; Q99836 transfection stap2-chuk 3 7 unspecified role unspecified role prey prey region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 9 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 unspecified role unspecified role prey prey region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association 3 C When expression vectors for FLAG-tagged MyD88 or His-tagged IKK- together with Myc-tagged STAP-2 were transfected into 293T cells and the cells were lysed, immunoprecipitated with an anti-Myc Ab, and immunoblotted with an anti-FLAG or anti-His Ab, the immunoprecipitates contained MyD88 and IKK- (Fig. 3, B and C), indicating that STAP-2 associated with MyD88 and IKK- in 293T cells. Q9UGK3; O15111; O14920 transfection stap2-chuk-1 4 7 unspecified role unspecified role prey prey 8 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association 3 D The Jurkat/STAP-2 cells were lysed, immunoprecipitated with an anti-Myc Ab and immunoblotted with an anti-IKK- Ab. As shown in Fig. 3D, the immunoprecipitates contained endogenous IKK- proteins. Q9UGK3 transfection stap2-ikbkb 1 9 unspecified role unspecified role bait bait 8 unspecified role unspecified role prey prey physical association physical association 3 E After treatment of THP-1 cells with PMA for 3 days, the cells were lysed and immunoprecipitated with a control Ab or anti-STAP-2 Ab. The immunoprecipitate with the anti-STAP-2 Ab contained IKK- ikbkb-myd88 3 9 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 unspecified role unspecified role prey prey region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 8 unspecified role unspecified role bait bait region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association 3 F we assessed the association of MyD88 with IKK- in the absence or presence of STAP-2. Expression vectors for FLAG-tagged MyD88 and Histagged IKK- together with or without Myc-tagged STAP-2 were transfected into 293T cells. The cells were lysed, immunoprecipitated with an anti-His Ab, and immunoblotted with an anti-FLAG or anti-His Ab. As shown in Fig. 3F, the association between MyD88 and IKK- was only detected in the presence of STAP-2. Therefore, STAP-2 associates with MyD88 and IKK-, and plays a role in forming a complex of these three molecules. The association between MyD88 and IKK-beta was only detected in the presence of STAP-2, indicating STAP-2 associates with MyD88 and IKK-beta, and plays a role in forming a complex of these three molecules. ikbkb-nfkbia-6 2 10 enzyme target enzyme target substrate substrate 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 8 enzyme enzyme unspecified role unspecified role region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined O14920:p.Lys44Met mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain K M Wed May 02 18:34:46 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys44met' Tue Jun 30 17:49:29 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys44Met' 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. phosphorylation phosphorylation reaction phosphorylation 4 C IKK--mediated incorporation of 32P into GST-IB as a substrate was also stimulated in the presence of STAP-2. GST-IkappaB is phosphorylated STAP-2 - This interaction was enhanced in the presence of increasing amounts of STAP-2. ikbkb-ikbkb-2 2 8 putative self putative self putative self putative self region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined O14920:p.Lys44Met mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain K M Wed May 02 14:33:46 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys44met' Tue Jun 30 18:14:17 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys44Met' 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. phosphorylation phosphorylation reaction phosphorylation true 4 C His-IKK-beta is autophosphorylated autophosphorylation of IKK- was markedly enhanced in the presence of higher amounts of STAP-2. STAP-2 - This interaction was enhanced in the presence of increasing amounts of STAP-2. myd88-stap2 3 9 unspecified role unspecified role prey prey 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 region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 6 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association 5 B the precipitate for GST-STAP-2 SH2 contained both MyD88 and IKK- Q99836; Q9UGK3 transfection The HS2-like domain of STAP-2 can bind MyD88. ikbkb-stap2-1 3 8 unspecified role unspecified role bait bait region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 9 unspecified role unspecified role prey prey 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 region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain physical association physical association 5 C the precipitate for GST-STAP-2 SH2 contained both MyD88 and IKK- Q9UGK3; O14920 transfection The SH2-like domain of STAP-2 can bind IKK-beta. ikbkb-stap2 3 9 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 8 unspecified role unspecified role bait bait region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association 5 D a deletion mutant of the SH2-like domain of STAP-2 failed to interact with IKK-, while full-length and deletion mutants of the PH or C-terminal domains of STAP-2 retained strong binding to IKK-. Therefore, the SH2-like domain of STAP-2 can bind to both MyD88 and IKK-. The SH2-like domain of STAP-2 can bind IKK-beta. stap2-ikbkb-1 5 8 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined n-terminal sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain c-teminal sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 9 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 physical association physical association 5 F 293T cells were transfected with GST-STAP-2 SH2 and/or a series of Myc-tagged IKK- deletion mutants (Fig. 5E). The transfectants were lysed, pulled down with glutathione-Sepharose beads (GSH bound), and immunoblotted with an anti-Myc Ab. As shown in Fig. 5F, the LZ domain of IKK- alone failed to interact with the SH2-like domain of STAP-2, while both the kinase and HLH domains of IKK- retained strong binding to STAP-2. STAP-2 interacts with the N-terminal and C-terminal domains of IKK-beta. stap2-myd88-1 5 6 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined n-terminal sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain c-teminal sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 9 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 physical association physical association 5 H Both the death domain and TIR domains of MyD88 interacted with the SH2-like domain of STAP-2. both the death domain and TIR domains of MyD88 interacted with the SH2-like domain of STAP-2.