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/ tornatore-2014-1 Cancer-selective targeting of the NF-κB survival pathway with GADD45β/MKK7 inhibitors. in vitro In vitro solid phase assay solid phase assay tag perox activity tag visualisation by peroxidase activity tag perox activity Tornatore L., Sandomenico A., Raimondo D., Low C., Rocci A., Tralau-Stewart C., Capece D., D'Andrea D., Bua M., Boyle E., van Duin M., Zoppoli P., Jaxa-Chamiec A., Thotakura A.K., Dyson J., Walker B.A., Leonardi A., Chambery A., Driessen C., Sonneveld P., Morgan G., Palumbo A., Tramontano A., Rahemtulla A., Ruvo M., Franzoso G. g.franzoso@imperial.ac.uk,menotti.ruvo@unina.it Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation tornatore-2014-5 Cancer-selective targeting of the NF-κB survival pathway with GADD45β/MKK7 inhibitors. in vitro In vitro cd circular dichroism CD CD cd predetermined predetermined participant predetermined Tornatore L., Sandomenico A., Raimondo D., Low C., Rocci A., Tralau-Stewart C., Capece D., D'Andrea D., Bua M., Boyle E., van Duin M., Zoppoli P., Jaxa-Chamiec A., Thotakura A.K., Dyson J., Walker B.A., Leonardi A., Chambery A., Driessen C., Sonneveld P., Morgan G., Palumbo A., Tramontano A., Rahemtulla A., Ruvo M., Franzoso G. g.franzoso@imperial.ac.uk,menotti.ruvo@unina.it Cellular based Kinase Assay results have not been included as the purity of the Kinase is not known. "Broad kinase-panel profiling" results have not been entered as the substrate(s) used are not known as they are not stated on the web-site of the International Centre for Kinase Profiling itself, nor the quoted references for methods used. Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation tornatore-2014-4 Cancer-selective targeting of the NF-κB survival pathway with GADD45β/MKK7 inhibitors. in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Tornatore L., Sandomenico A., Raimondo D., Low C., Rocci A., Tralau-Stewart C., Capece D., D'Andrea D., Bua M., Boyle E., van Duin M., Zoppoli P., Jaxa-Chamiec A., Thotakura A.K., Dyson J., Walker B.A., Leonardi A., Chambery A., Driessen C., Sonneveld P., Morgan G., Palumbo A., Tramontano A., Rahemtulla A., Ruvo M., Franzoso G. Tryptophan fluorescence quenching g.franzoso@imperial.ac.uk,menotti.ruvo@unina.it Cellular based Kinase Assay results have not been included as the purity of the Kinase is not known. Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation tornatore-2014-6 Cancer-selective targeting of the NF-κB survival pathway with GADD45β/MKK7 inhibitors. in vitro In vitro ms interact detect detection by mass spectrometry ms interact detect predetermined predetermined participant predetermined Tornatore L., Sandomenico A., Raimondo D., Low C., Rocci A., Tralau-Stewart C., Capece D., D'Andrea D., Bua M., Boyle E., van Duin M., Zoppoli P., Jaxa-Chamiec A., Thotakura A.K., Dyson J., Walker B.A., Leonardi A., Chambery A., Driessen C., Sonneveld P., Morgan G., Palumbo A., Tramontano A., Rahemtulla A., Ruvo M., Franzoso G. g.franzoso@imperial.ac.uk,menotti.ruvo@unina.it Cellular based Kinase Assay results have not been included as the purity of the Kinase is not known. "Broad kinase-panel profiling" results have not been entered as the substrate(s) used are not known as they are not stated on the web-site of the International Centre for Kinase Profiling itself, nor the quoted references for methods used. Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation tornatore-2014-2 Cancer-selective targeting of the NF-κB survival pathway with GADD45β/MKK7 inhibitors. human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Tornatore L., Sandomenico A., Raimondo D., Low C., Rocci A., Tralau-Stewart C., Capece D., D'Andrea D., Bua M., Boyle E., van Duin M., Zoppoli P., Jaxa-Chamiec A., Thotakura A.K., Dyson J., Walker B.A., Leonardi A., Chambery A., Driessen C., Sonneveld P., Morgan G., Palumbo A., Tramontano A., Rahemtulla A., Ruvo M., Franzoso G. g.franzoso@imperial.ac.uk,menotti.ruvo@unina.it Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation tornatore-2014-3 Cancer-selective targeting of the NF-κB survival pathway with GADD45β/MKK7 inhibitors. in vitro In vitro fluorescence spectr classical fluorescence spectroscopy fluorescence spectr predetermined predetermined participant predetermined Tornatore L., Sandomenico A., Raimondo D., Low C., Rocci A., Tralau-Stewart C., Capece D., D'Andrea D., Bua M., Boyle E., van Duin M., Zoppoli P., Jaxa-Chamiec A., Thotakura A.K., Dyson J., Walker B.A., Leonardi A., Chambery A., Driessen C., Sonneveld P., Morgan G., Palumbo A., Tramontano A., Rahemtulla A., Ruvo M., Franzoso G. Tryptophan fluorescence quenching g.franzoso@imperial.ac.uk,menotti.ruvo@unina.it Cellular based Kinase Assay results have not been included as the purity of the Kinase is not known. Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation ga45b_human Growth arrest and DNA damage-inducible protein GADD45 beta Myeloid differentiation primary response protein MyD118 Negative growth regulatory protein MyD118 GADD45B MYD118 protein protein human Homo sapiens Human MTLEELVACDNAAQKMQTVTAAVEELLVAAQRQDRLTVGVYESAKLMNVDPDSVVLCLLAIDEEEEDDIALQIHFTLIQSFCCDNDINIVRVSGMQRLAQLLGEPAETQGTTEARDLHCLLVTNPHTDAWKSHGLVEVASYCEESRGNNQWVPYISLQER mp2k7_human Dual specificity mitogen-activated protein kinase kinase 7 MAPK/ERK kinase 7 JNK-activating kinase 2 c-Jun N-terminal kinase kinase 2 MAP2K7 JNKK2 MKK7 PRKMK7 MEK7 Stress-activated protein kinase kinase 4 SKK4 protein protein human Homo sapiens Human MAASSLEQKLSRLEAKLKQENREARRRIDLNLDISPQRPRPTLQLPLANDGGSRSPSSESSPQHPTPPARPRHMLGLPSTLFTPRSMESIEIDQKLQEIMKQTGYLTIGGQRYQAEINDLENLGEMGSGTCGQVWKMRFRKTGHVIAVKQMRRSGNKEENKRILMDLDVVLKSHDCPYIVQCFGTFITNTDVFIAMELMGTCAEKLKKRMQGPIPERILGKMTVAIVKALYYLKEKHGVIHRDVKPSNILLDERGQIKLCDFGISGRLVDSKAKTRSAGCAAYMAPERIDPPDPTKPDYDIRADVWSLGISLVELATGQFPYKNCKTDFEVLTKVLQEEPPLLPGHMGFSGDFQSFVKDCLTKDHRKRPKYNKLLEHSFIKRYETLEVDVASWFKDVMAKTESPRTSGVLSQPHLPFFR (dtp3)-tyr,arg,phe (DTP3)-D_tripeptide_ac_tyr-arg-phe peptide peptide polypeptide oligopeptide oligopeptide polypeptide chemical synthesis Chemical synthesis (Chemical synthesis) ac-ltp1_tyr-asp-his-phe Ac-LTP1-L-stereoisomer peptide peptide polypeptide oligopeptide oligopeptide polypeptide chemical synthesis Chemical synthesis (Chemical synthesis) ac-ltp2_tyr-glu-arg-phe Ac-LTP2-L-stereoisomer peptide peptide polypeptide oligopeptide oligopeptide polypeptide chemical synthesis Chemical synthesis (Chemical synthesis) ac-dtp1_tyr-asp-his-phe Ac-DTP1-D-stereoisomer peptide peptide polypeptide oligopeptide oligopeptide polypeptide chemical synthesis Chemical synthesis (Chemical synthesis) ac-dtp2_tyr-glu-arg-phe Ac-DTP2-D-stereoisomer peptide peptide polypeptide oligopeptide oligopeptide polypeptide chemical synthesis Chemical synthesis (Chemical synthesis) map2k7-ac_dtp2 1765209 n/a 1765215 unspecified role unspecified role prey prey region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) n/a 1765216 unspecified role unspecified role bait bait n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) n/a 1765217 competitor competitor competitor prey prey n_terminus ac1res monoacetylated residue Acetyl Acetylation (N terminus, N epsilon of Lysine, O of Serine) (Ac) Acetylation Ac1Res n-term range n-terminal range n-term range n-term range n-terminal range n-term range false chemical synthesis Chemical synthesis (Chemical synthesis) physical association physical association false false false 1765209 S5D map2k7-gadd45b 1765209 n/a 1765216 unspecified role unspecified role bait bait n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) n/a 1765215 unspecified role unspecified role prey prey n_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false Fig 3, S4 A/B map2k7-ac_ltp1-1 1765209 n/a 1765218 competitor competitor competitor prey prey n_terminus ac1res monoacetylated residue Acetyl Acetylation (N terminus, N epsilon of Lysine, O of Serine) (Ac) Acetylation Ac1Res n-term range n-terminal range n-term range n-term range n-terminal range n-term range false chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1765215 unspecified role unspecified role prey prey n_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) n/a 1765216 unspecified role unspecified role bait bait n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) physical association physical association false false false 1765209 Fig 3i, The two L-tetrapeptides lost activity after incubation with human serum, unlike the two D-enantiomers. preincubation with human serum map2k7-ac_ltp1-2 1765209 n/a 1765219 competitor competitor competitor prey prey n_terminus ac1res monoacetylated residue Acetyl Acetylation (N terminus, N epsilon of Lysine, O of Serine) (Ac) Acetylation Ac1Res n-term range n-terminal range n-term range n-term range n-terminal range n-term range false chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1765215 unspecified role unspecified role prey prey region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) n/a 1765216 unspecified role unspecified role bait bait n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) physical association physical association false false false 1765209 Fig 3ii The two L-tetrapeptides lost activity after incubation with human serum, unlike the two D-enantiomers. preincubation with human serum map2k7-ac_ltp1-3 1765209 n/a 1765220 competitor competitor competitor prey prey n_terminus ac1res monoacetylated residue Acetyl Acetylation (N terminus, N epsilon of Lysine, O of Serine) (Ac) Acetylation Ac1Res n-term range n-terminal range n-term range n-term range n-terminal range n-term range false chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1765215 unspecified role unspecified role prey prey n_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false ecoli Escherichia coli (strain K12) n/a 1765216 unspecified role unspecified role bait bait n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) physical association physical association false false false 1765209 Fig 3iii, S4 A/B The two L-tetrapeptides lost activity after incubation with human serum, unlike the two D-enantiomers. Replacement of the N-terminal acetyl group of Ac-DTP1 and Ac-DTP2 with a benzyloxycarbonyl [Z] group, resulted in no loss of activity. map2k7-ac_ltp2 1765209 n/a 1765215 unspecified role unspecified role prey prey region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) n/a 1765221 competitor competitor competitor prey prey n_terminus ac1res monoacetylated residue Acetyl Acetylation (N terminus, N epsilon of Lysine, O of Serine) (Ac) Acetylation Ac1Res n-term range n-terminal range n-term range n-term range n-terminal range n-term range false chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1765216 unspecified role unspecified role bait bait n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) physical association physical association false false false 1765209 Fig 3iv, S4 A/B The two L-tetrapeptides lost activity after incubation with human serum, unlike the two D-enantiomers. Replacement of the N-terminal acetyl group of Ac-DTP1 and Ac-DTP2 with a benzyloxycarbonyl [Z] group, resulted in no loss of activity. gadd45b-map2k7 1765210 n/a 1765215 unspecified role unspecified role neutral component neutral component n_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) n/a 1765216 unspecified role unspecified role neutral component neutral component n_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false kinase_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) direct interaction direct interaction false false false Supp fig 5I _dtp3__tyr_arg_phe-gadd45b 1765210 n/a 1765215 unspecified role unspecified role neutral component neutral component n_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) n/a 1765216 unspecified role unspecified role neutral component neutral component n_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false kinase_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 1765217 unspecified role unspecified role neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) direct interaction direct interaction false false false Supp fig 5Ii DTP3 can cause a significant conformational change of the structure of the GADD45beta/MKK7complex, as well as that of the isolated MKK7 protein. _dtp3__tyr_arg_phe-gadd45b-1 1765210 n/a 1765216 unspecified role unspecified role neutral component neutral component n_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false kinase_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 1765217 unspecified role unspecified role neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) direct interaction direct interaction false false false Supp fig 5Iii DTP3 can cause a significant conformational change of the structure of the GADD45beta/MKK7complex, as well as that of the isolated MKK7 protein. _dtp3__tyr_arg_phe-map2k7 1765211 n/a 1765217 unspecified role unspecified role bait bait n_terminus ac1res monoacetylated residue Acetyl Acetylation (N terminus, N epsilon of Lysine, O of Serine) (Ac) Acetylation Ac1Res n-term range n-terminal range n-term range n-term range n-terminal range n-term range false chemical synthesis Chemical synthesis (Chemical synthesis) Stoichiometry: 1.0 n/a 1765216 unspecified role unspecified role prey prey n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false human-kms12 KMS12 Myeloma cell line kms12 KMS12 Myeloma cell line Stoichiometry: 1.0 physical association physical association false false false Fig 5E _dtp3__tyr_arg_phe-map2k7-1 1765211 n/a 1765217 unspecified role unspecified role bait bait n_terminus ac1res monoacetylated residue Acetyl Acetylation (N terminus, N epsilon of Lysine, O of Serine) (Ac) Acetylation Ac1Res n-term range n-terminal range n-term range n-term range n-terminal range n-term range false chemical synthesis Chemical synthesis (Chemical synthesis) Stoichiometry: 1.0 n/a 1765216 unspecified role unspecified role prey prey n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false human-u266 Homo sapiens myeloma cell line U-266 u266 Human myeloma cell line U-266 confirmed as human with IEF of AST, MDH, NP established from the peripheral blood of a 53-year-old man with IgE-secreting myeloma (refractory, terminal) in 1968; cells were described to produce IgE lambda; cells express mRNA for BCL2 gene Stoichiometry: 1.0 physical association physical association false false false Fig 5Ei _dtp3__tyr_arg_phe-gadd45b-2 1765212 n/a 1765216 unspecified role unspecified role neutral component neutral component kinase_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) n/a 1765217 unspecified role unspecified role neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) direct interaction direct interaction false false false Supp fig 5Iii DTP3 can cause a significant conformational change of the structure of the GADD45beta/MKK7complex, as well as that of the isolated MKK7 protein. map2k7-gadd45b-1 1765213 n/a 1765216 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 false n/a 1765215 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false DTP3 ac-DTP2 Fig 3C, Supp fig 5E ac-DTP1 dtp3-map2k7 1765214 n/a 1765217 unspecified role unspecified role neutral component neutral component n_terminus ac1res monoacetylated residue Acetyl Acetylation (N terminus, N epsilon of Lysine, O of Serine) (Ac) Acetylation Ac1Res n-term range n-terminal range n-term range n-term range n-terminal range n-term range false chemical synthesis Chemical synthesis (Chemical synthesis) Stoichiometry: 1.0 n/a 1765216 unspecified role unspecified role neutral component neutral component n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false ecoli Escherichia coli (strain K12) Stoichiometry: 1.0 direct interaction direct interaction false false false 1765214 Fig 5B, Supp 5K