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/ Overexpression of Tissue Transglutaminase Leads to Constitutive Activation of Nuclear Factor-{kappa}B in Cancer Cells: Delineation of a Novel Pathway. Overexpression of Tissue Transglutaminase Leads to Constitutive Activation of Nuclear Factor-{kappa}B in Cancer Cells: Delineation of a Novel Pathway. Cancer Res. (0008-5472) 2006 imex curation Mann AP., Verma A., Sethi G., Manavathi B., Wang H., Fok JY., Kunnumakkara AB., Kumar R., Aggarwal BB., Mehta K. Cancer - Interactions investigated in the context of cancer kmehta@mdanderson.org 13-Nov-2007: Contacted by JYOTI. orchard Only protein-protein interactions human-mdapanc28 Homo sapiens pancreatic adenocarcinoma cell line mdapanc28 Human pancreatic adenocarcinoma cell line The cell line expresses trypsin and ribonuclease RNA, which are considered to be acinar cell markers, and carbonic anhydrase II (CAII), which is considered to be a duct-cell markers. The histological appearance of xenografts in nude mice was similar to that of the tumor from which the cell line was established. The chromosome number varied between 46 and 60. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Mann AP., Verma A., Sethi G., Manavathi B., Wang H., Fok JY., Kunnumakkara AB., Kumar R., Aggarwal BB., Mehta K. Cancer - Interactions investigated in the context of cancer kmehta@mdanderson.org Accepted 2007-NOV-2 by ORCHARD. Cancer Res. (0008-5472) 2006 orchard Only protein-protein interactions imex curation Overexpression of Tissue Transglutaminase Leads to Constitutive Activation of Nuclear Factor-{kappa}B in Cancer Cells: Delineation of a Novel Pathway. Overexpression of Tissue Transglutaminase Leads to Constitutive Activation of Nuclear Factor-{kappa}B in Cancer Cells: Delineation of a Novel Pathway. Cancer Res. (0008-5472) 2006 imex curation Mann AP., Verma A., Sethi G., Manavathi B., Wang H., Fok JY., Kunnumakkara AB., Kumar R., Aggarwal BB., Mehta K. Cancer - Interactions investigated in the context of cancer kmehta@mdanderson.org 13-Nov-2007: Contacted by JYOTI. orchard Only protein-protein interactions human-mcf7 Homo sapiens mammary carcinoma cells mcf7 mammary carcinoma cells anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Mann AP., Verma A., Sethi G., Manavathi B., Wang H., Fok JY., Kunnumakkara AB., Kumar R., Aggarwal BB., Mehta K. Cancer - Interactions investigated in the context of cancer kmehta@mdanderson.org Accepted 2007-NOV-2 by ORCHARD. 13-Nov-2007: Contacted by JYOTI. Cancer Res. (0008-5472) 2006 orchard Only protein-protein interactions imex curation Overexpression of Tissue Transglutaminase Leads to Constitutive Activation of Nuclear Factor-{kappa}B in Cancer Cells: Delineation of a Novel Pathway. Overexpression of Tissue Transglutaminase Leads to Constitutive Activation of Nuclear Factor-{kappa}B in Cancer Cells: Delineation of a Novel Pathway. Cancer Res. (0008-5472) 2006 imex curation Mann AP., Verma A., Sethi G., Manavathi B., Wang H., Fok JY., Kunnumakkara AB., Kumar R., Aggarwal BB., Mehta K. Cancer - Interactions investigated in the context of cancer kmehta@mdanderson.org 13-Nov-2007: Contacted by JYOTI. orchard Only protein-protein interactions human-bxpc3 Homo sapiens primary pancreatic adenocarcinoma cell line bxpc3 BxPC-3 Human primary pancreatic adenocarcinoma cell line Derived from a 61 year old female with a primary adenocarcinoma of the pancreas. BxPC-3 cells produce mucin and the tumour in a nude mouse is moderately well to poorly differentiated like the primary adenocarcinoma anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Mann AP., Verma A., Sethi G., Manavathi B., Wang H., Fok JY., Kunnumakkara AB., Kumar R., Aggarwal BB., Mehta K. Cancer - Interactions investigated in the context of cancer kmehta@mdanderson.org Accepted 2007-NOV-2 by ORCHARD. 13-Nov-2007: Contacted by JYOTI. Cancer Res. (0008-5472) 2006 orchard Only protein-protein interactions imex curation Overexpression of Tissue Transglutaminase Leads to Constitutive Activation of Nuclear Factor-{kappa}B in Cancer Cells: Delineation of a Novel Pathway. Overexpression of Tissue Transglutaminase Leads to Constitutive Activation of Nuclear Factor-{kappa}B in Cancer Cells: Delineation of a Novel Pathway. Cancer Res. (0008-5472) 2006 imex curation Mann AP., Verma A., Sethi G., Manavathi B., Wang H., Fok JY., Kunnumakkara AB., Kumar R., Aggarwal BB., Mehta K. Cancer - Interactions investigated in the context of cancer kmehta@mdanderson.org 13-Nov-2007: Contacted by JYOTI. orchard Only protein-protein interactions human-mdapanc28 Homo sapiens pancreatic adenocarcinoma cell line mdapanc28 Human pancreatic adenocarcinoma cell line The cell line expresses trypsin and ribonuclease RNA, which are considered to be acinar cell markers, and carbonic anhydrase II (CAII), which is considered to be a duct-cell markers. The histological appearance of xenografts in nude mice was similar to that of the tumor from which the cell line was established. The chromosome number varied between 46 and 60. confocal microscopy confocal microscopy predetermined predetermined participant predetermined Mann AP., Verma A., Sethi G., Manavathi B., Wang H., Fok JY., Kunnumakkara AB., Kumar R., Aggarwal BB., Mehta K. Cancer - Interactions investigated in the context of cancer kmehta@mdanderson.org Accepted 2007-NOV-2 by ORCHARD. 13-Nov-2007: Contacted by JYOTI. Cancer Res. (0008-5472) 2006 orchard Only protein-protein interactions imex curation 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 tf65_human Transcription factor p65 Nuclear factor NF-kappa-B p65 subunit RELA NFKB3 Nuclear factor of kappa light polypeptide gene enhancer in B-cells 3 protein protein human Homo sapiens Human MDELFPLIFPAEPAQASGPYVEIIEQPKQRGMRFRYKCEGRSAGSIPGERSTDTTKTHPTIKINGYTGPGTVRISLVTKDPPHRPHPHELVGKDCRDGFYEAELCPDRCIHSFQNLGIQCVKKRDLEQAISQRIQTNNNPFQVPIEEQRGDYDLNAVRLCFQVTVRDPSGRPLRLPPVLSHPIFDNRAPNTAELKICRVNRNSGSCLGGDEIFLLCDKVQKEDIEVYFTGPGWEARGSFSQADVHRQVAIVFRTPPYADPSLQAPVRVSMQLRRPSDRELSEPMEFQYLPDTDDRHRIEEKRKRTYETFKSIMKKSPFSGPTDPRPPPRRIAVPSRSSASVPKPAPQPYPFTSSLSTINYDEFPTMVFPSGQISQASALAPAPPQVLPQAPAPAPAPAMVSALAQAPAPVPVLAPGPPQAVAPPAPKPTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNSEFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGDEDFSSIADMDFSALLSQISS DIP protein Q04206 original sequence version: 170 DIP protein Q04206 original sequence version: 173 DIP protein Q04206 original sequence version: 99 DIP protein Q04206-1 original sequence version: 99 DIP protein Q04206 original sequence version: 168 DIP protein Q04206 original sequence version: 128 DIP protein Q04206 original sequence version: 131 DIP protein Q04206 original sequence version: 153 DIP protein Q04206 original sequence version: 185 DIP protein Q04206 original sequence version: 181 DIP protein Q04206 original sequence version: 183 8147A6AF9F2445C6 tgm2_human Protein-glutamine gamma-glutamyltransferase 2 TGM2 Tissue transglutaminase Transglutaminase C Transglutaminase-2 Transglutaminase H Erythrocyte transglutaminase Heart G alpha(h) Isopeptidase TGM2 Protein G alpha(h) Protein-glutamine deamidase TGM2 Protein-glutamine dopaminyltransferase TGM2 Protein-glutamine histaminyltransferase TGM2 Protein-glutamine noradrenalinyltransferase TGM2 Protein-glutamine serotonyltransferase TGM2 Transglutaminase II protein protein human Homo sapiens Human MAEELVLERCDLELETNGRDHHTADLCREKLVVRRGQPFWLTLHFEGRNYEASVDSLTFSVVTGPAPSQEAGTKARFPLRDAVEEGDWTATVVDQQDCTLSLQLTTPANAPIGLYRLSLEASTGYQGSSFVLGHFILLFNAWCPADAVYLDSEEERQEYVLTQQGFIYQGSAKFIKNIPWNFGQFEDGILDICLILLDVNPKFLKNAGRDCSRRSSPVYVGRVVSGMVNCNDDQGVLLGRWDNNYGDGVSPMSWIGSVDILRRWKNHGCQRVKYGQCWVFAAVACTVLRCLGIPTRVVTNYNSAHDQNSNLLIEYFRNEFGEIQGDKSEMIWNFHCWVESWMTRPDLQPGYEGWQALDPTPQEKSEGTYCCGPVPVRAIKEGDLSTKYDAPFVFAEVNADVVDWIQQDDGSVHKSINRSLIVGLKISTKSVGRDEREDITHTYKYPEGSSEEREAFTRANHLNKLAEKEETGMAMRIRVGQSMNMGSDFDVFAHITNNTAEEYVCRLLLCARTVSYNGILGPECGTKYLLNLNLEPFSEKSVPLCILYEKYRDCLTESNLIKVRALLVEPVINSYLLAERDLYLENPEIKIRILGEPKQKRKLVAEVSLQNPLPVALEGCTFTVEGAGLTEEQKTVEIPDPVEAGEEVKVRMDLLPLHMGLHKLVVNFESDKLKAVKGFRNVIIGPA DIP protein P21980 original sequence version: 125 7DA33FF335DE7B37 nfkbia-rela-1 1 6 unspecified role unspecified role prey prey 7 unspecified role unspecified role prey prey 5 unspecified role unspecified role bait bait association association 4 A Immunoprecipitation of Panc28 cytoplasmic extracts with anti-InBa antibody effectively pulled down the TG2 protein in addition to the InBa and p65 proteins, suggesting that TG2 forms a part of the complex between p65/p50 and InBa A23187 - Treatment with the calcium ionophore, A23187, (which causes activation of TG2), enhanced this interaction. nfkbia-rela-2 1 6 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait physical association physical association 4 C We confirmed this association by immunoprecipitating TG2 and showing the presence of p65 and InBa in the immune complex Note that the blot in Fig. 4 C only shows that p65 is coimmunoprecipitated with TG2 and IkappaBalpha is not shown to coimmunoprecipitate. nfkbia-rela-3 2 5 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey physical association physical association 4 A Unlike in the MDAPanc28 lysates, IkappaBalpha did not also coimmunoprecipitate TG2. tgm2-rela 3 7 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey physical association physical association 4 C rela-tgm2 4 6 unspecified role unspecified role neutral component neutral component 7 unspecified role unspecified role neutral component neutral component colocalization colocalization confocal microscopy data supported the colocalization of TG2 with p65 in the cytoplasm (Fig. 4B). The association between TG2 and p65 further increased in response to treatment with A23187, which led to increased translocation of the p65/TG2 complex in the nucleus 4 B A23187 - Treatment with the calcium ionophore, A23187, which activates TG2, enahnced complex formation and increased translocation of the complex to the nucleus. nfkb1-nfkb1-2 1 5 unspecified role unspecified role prey prey 5 unspecified role unspecified role bait bait physical association physical association 4 D Immunoprecipitation of a cytoplasmic extract from TG2-rich Panc28 cells with an anti-InBa antibody revealed the presence of a 66-kDa dimeric InBa band (Fig. 4D, left) in addition to the 33-kDa monomeric InBa band. Treatment of cells with A23187 further augmented the formation of dimeric bands and resulted in the appearance of another polymeric InBa band. A23187 - Treatment with the calcium ionophore (A23187), which activates TG2, augmented dimer formation and results in the apppearance of another polymeric IKappaBalpha band. The dimeric form of IkappaBalpha was also seen when IKappaBalpha immunoprecipitates were labeled with BPA and probed with HRP-conjugated streptavidin. nfkb1-nfkb1-3 1 5 unspecified role unspecified role bait bait 5 unspecified role unspecified role prey prey physical association physical association 4 D Immunoprecipitation of a cytoplasmic extract from TG2-rich Panc28 cells with an anti-InBa antibody revealed the presence of a 66-kDa dimeric InBa band (Fig. 4D, left) in addition to the 33-kDa monomeric InBa band. Treatment of cells with A23187 further augmented the formation of dimeric bands and resulted in the appearance of another polymeric InBa band. A23187 - Treatment with the calcium ionophore (A23187), which activates TG2, augmented dimer formation and results in the apppearance of another polymeric IKappaBalpha band. This polymeric form of IkappaBalpha is of higher order than the dimeric form also seen, as A23187 treatment resulted in the appearance of another polymeric band. The polymeric form of IkappaBalpha was also seen when IKappaBalpha immunoprecipitates were labeled with BPA and probed with HRP-conjugated streptavidin.