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/ chen-2014a-1 Small GTPase RBJ Mediates Nuclear Entrapment of MEK1/MEK2 in Tumor Progression. in vitro In vitro pull down pull down autoradiography autoradiography Chen T., Yang M., Yu Z., Tang S., Wang C., Zhu X., Guo J., Li N., Zhang W., Hou J., Liu H., Han C., Liu Q., Gu Y., Qian C., Wan T., Cui L., Zhu M., Zheng W., Cao X. Cancer - Interactions investigated in the context of cancer Results obtained using the anti-ERK1/2 and MEK1/2 antibody have not been included due to lack of antibody specificity - as in Figure 4. The origins of HA/GST-RBJ and mutants, GST-KRAS,GST-Rab5A and Recombinant HSC70 cannot be found, and the authors did not reply to a request for info. The quoted 'Wang et al., 2009 reference does not mention these constructs;as a consequence results from several figures cannot be included e.g.Figures 4, 6, Supp. Figure 2. Samples were dotted onto nitrocellulose membranes and autoradiographed. caoxt@immunol.org Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation chen-2014a-3 Small GTPase RBJ Mediates Nuclear Entrapment of MEK1/MEK2 in Tumor Progression. in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Chen T., Yang M., Yu Z., Tang S., Wang C., Zhu X., Guo J., Li N., Zhang W., Hou J., Liu H., Han C., Liu Q., Gu Y., Qian C., Wan T., Cui L., Zhu M., Zheng W., Cao X. Cancer - Interactions investigated in the context of cancer Results obtained using the anti-ERK1/2 and MEK1/2 antibody have not been included due to lack of antibody specificity - as in Figure 4. The origins of HA/GST-RBJ and mutants, GST-KRAS,GST-Rab5A and Recombinant HSC70 cannot be found, and the authors did not reply to a request for info. The quoted 'Wang et al., 2009 reference does not mention these constructs;as a consequence results from several figures cannot be included e.g.Figures 4, 6, Supp. Figure 2. caoxt@immunol.org Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation chen-2014a-4 Small GTPase RBJ Mediates Nuclear Entrapment of MEK1/MEK2 in Tumor Progression. in vitro In vitro pull down pull down tag visualisation tag visualisation Chen T., Yang M., Yu Z., Tang S., Wang C., Zhu X., Guo J., Li N., Zhang W., Hou J., Liu H., Han C., Liu Q., Gu Y., Qian C., Wan T., Cui L., Zhu M., Zheng W., Cao X. Cancer - Interactions investigated in the context of cancer Results obtained using the anti-ERK1/2 and MEK1/2 antibody have not been included due to lack of antibody specificity - as in Figure 4. The origins of HA/GST-RBJ and mutants, GST-KRAS,GST-Rab5A and Recombinant HSC70 cannot be found, and the authors did not reply to a request for info. The quoted 'Wang et al., 2009 reference does not mention these constructs;as a consequence results from several figures cannot be included e.g.Figures 4, 6, Supp. Figure 2. Samples were prepared for scintillation counting. caoxt@immunol.org Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation chen-2014a-2 Small GTPase RBJ Mediates Nuclear Entrapment of MEK1/MEK2 in Tumor Progression. in vitro In vitro gtpase assay gtpase assay GTPase gtp hydrolisis gtp hydrolisis GTPase autoradiography autoradiography Chen T., Yang M., Yu Z., Tang S., Wang C., Zhu X., Guo J., Li N., Zhang W., Hou J., Liu H., Han C., Liu Q., Gu Y., Qian C., Wan T., Cui L., Zhu M., Zheng W., Cao X. Cancer - Interactions investigated in the context of cancer Results obtained using the anti-ERK1/2 and MEK1/2 antibody have not been included due to lack of antibody specificity - as in Figure 4. The origins of HA/GST-RBJ and mutants, GST-KRAS,GST-Rab5A and Recombinant HSC70 cannot be found, and the authors did not reply to a request for info. The quoted 'Wang et al., 2009 reference does not mention these constructs;as a consequence results from several figures cannot be included e.g.Figures 4, 6, Supp. Figure 2. The extent of GTP hydrolysis was assessed by measuring the remaining radioactivity- for GTP-bound protein. caoxt@immunol.org Cancer Cell (1535-6108) 2014 Only protein-protein interactions imex curation gtp guanosine triphosphate - GTP 5'-GTP GTP Guanosine 5'-triphosphate guanosine 5'-triphosphoric acid guanosine triphosphate GUANOSINE-5'-TRIPHOSPHATE H4gtp guanosine 5'-(tetrahydrogen triphosphate) small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) InChI=1/C10H16N5O14P3/c11-10-13-7-4(8(18)14-10)12-2-15(7)9-6(17)5(16)3(27-9)1-26-31(22,23)29-32(24,25)28-30(19,20)21/h2-3,5-6,9,16-17H,1H2,(H,22,23)(H,24,25)(H2,19,20,21)(H3,11,13,14,18)/t3-,5-,6-,9-/m1/s1/f/h14,19-20,22,24H,11H2 djc27_mouse DnaJ homolog subfamily C member 27 Dnajc27 Rabj Rbj Rab and DnaJ domain-containing protein protein protein mouse Mus musculus Mouse METNVPKRKEPAKSLRIKVISMGNAEVGKSCIIKRYCEKRFVSKYLATIGIDYGVTKVQVRDREIKVNIFDMAGHPFFFEVRNEFYKDTQGVILVYDVGQKDSFDALDSWLAEMKQELGPHGNMDNIVFVVCANKIDCSKHRCIDESEGRLWAESKGFLYFETSAQTGEGINEMFQTFYLSIVDLCENGGKRPTASSSASFTKEQADTIRRIRNSKDSWEMLGVRPGASREEVNKAYRKLAVLLHPDKCVAPGSEDAFKAVVNARTALLKNIK gdp guanosine 5-diphosphate small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) guanosine 5'-[gamma-thio]triphosphate GTP-gamma-S small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) mp2k1_mouse Dual specificity mitogen-activated protein kinase kinase 1 Map2k1 Mek1 Prkmk1 ERK activator kinase 1 MAPK/ERK kinase 1 protein protein mouse Mus musculus Mouse MPKKKPTPIQLNPAPDGSAVNGTSSAETNLEALQKKLEELELDEQQRKRLEAFLTQKQKVGELKDDDFEKISELGAGNGGVVFKVSHKPSGLVMARKLIHLEIKPAIRNQIIRELQVLHECNSPYIVGFYGAFYSDGEISICMEHMDGGSLDQVLKKAGRIPEQILGKVSIAVIKGLTYLREKHKIMHRDVKPSNILVNSRGEIKLCDFGVSGQLIDSMANSFVGTRSYMSPERLQGTHYSVQSDIWSMGLSLVEMAVGRYPIPPPDAKELELLFGCHVEGDAAETPPRPRTPGRPLSSYGMDSRPPMAIFELLDYIVNEPPPKLPSGVFSLEFQDFVNKCLIKNPAERADLKQLMVHAFIKRSDAEEVDFAGWLCSTIGLNQPSTPTHAASI mk01_mouse Mitogen-activated protein kinase 1 Mapk1 Erk2 Mapk Prkm1 Extracellular signal-regulated kinase 2 Mitogen-activated protein kinase 2 MAP kinase isoform p42 ERT1 protein protein mouse Mus musculus Mouse MAAAAAAGPEMVRGQVFDVGPRYTNLSYIGEGAYGMVCSAYDNLNKVRVAIKKISPFEHQTYCQRTLREIKILLRFRHENIIGINDIIRAPTIEQMKDVYIVQDLMETDLYKLLKTQHLSNDHICYFLYQILRGLKYIHSANVLHRDLKPSNLLLNTTCDLKICDFGLARVADPDHDHTGFLTEYVATRWYRAPEIMLNSKGYTKSIDIWSVGCILAEMLSNRPIFPGKHYLDQLNHILGILGSPSQEDLNCIINLKARNYLLSLPHKNKVPWNRLFPNADSKALDLLDKMLTFNPHKRIEVEQALAHPYLEQYYDPSDEPIAEAPFKFDMELDDLPKEKLKELIFEETARFQPGYRS rask_human GTPase KRas KRAS RASK2 KRAS2 K-Ras 2 Ki-Ras c-K-ras c-Ki-ras protein protein human Homo sapiens Human MTEYKLVVVGAGGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVVIDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHHYREQIKRVKDSEDVPMVLVGNKCDLPSRTVDTKQAQDLARSYGIPFIETSAKTRQRVEDAFYTLVREIRQYRLKKISKEEKTPGCVKIKKCIIM dnajc27-gtp 1546711 n/a 1546715 unspecified role unspecified role prey prey region 32p radiolabel 32p radiolabel P32 32P 32P P32 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1546716 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 small_gtp_binding_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Q8CFP6:p.Ser30Asn mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false 2A dnajc27-gdp 1546711 n/a 1546717 competitor competitor competitor prey prey chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1546715 unspecified role unspecified role prey prey region 32p radiolabel 32p radiolabel P32 32P 32P P32 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1546716 unspecified role unspecified role bait bait Q8CFP6:p.Ser30Asn mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false 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) direct interaction direct interaction false false false 2Ai dnajc27-gdp-1 1546711 n/a 1546715 unspecified role unspecified role prey prey region 32p radiolabel 32p radiolabel P32 32P 32P P32 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1546716 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 Q8CFP6:p.Ser30Asn mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1546718 competitor competitor competitor prey prey chemical synthesis Chemical synthesis (Chemical synthesis) direct interaction direct interaction false false false 2Aii dnajc27-gtp-4 1546711 n/a 1546715 unspecified role unspecified role prey prey region 32p radiolabel 32p radiolabel P32 32P 32P P32 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1546716 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 small_gtp_binding_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Q8CFP6:p.Ser30Asn mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1546715 competitor competitor competitor prey prey chemical synthesis Chemical synthesis (Chemical synthesis) direct interaction direct interaction false false false 2A... dnajc27-gtp-3 1546712 n/a 1546719 unspecified role unspecified role prey prey c_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false ecoli Escherichia coli (strain K12) n/a 1546716 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 small_gtp_binding_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false Q8CFP6:p.Ser30Asn mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false 4B/F dnajc27-map2k1 1546712 n/a 1546716 unspecified role unspecified role bait bait n_terminal_region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain false 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 1546720 unspecified role unspecified role prey prey c_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false 4B/F,6H dnajc27-gdp-2 1546712 n/a 1546719 unspecified role unspecified role prey prey c_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false ecoli Escherichia coli (strain K12) n/a 1546716 unspecified role unspecified role bait bait small_gtp_binding_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 Q8CFP6:p.Ser30Asn mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) direct interaction direct interaction false false false 4F In the presence of excessive GDP the interaction was reduced. The authors conclude that RBJ interacted with MEK1 in its GTP-bound status. dnajc27-map2k1-2 1546712 n/a 1546716 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 1546719 unspecified role unspecified role prey prey c_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false ecoli Escherichia coli (strain K12) n/a 1546719 competitor competitor competitor prey prey nes_region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false direct interaction direct interaction false false false 6H dnajc27-gdp-3 1546713 n/a 1546718 competitor competitor competitor prey prey chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1546715 unspecified role unspecified role prey prey region 32p radiolabel 32p radiolabel P32 32P 32P P32 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1546716 unspecified role unspecified role bait bait Q8CFP6:p.Ser30Asn mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false 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) direct interaction direct interaction false false false Supp. 2C dnajc27-gdp-3 1546713 n/a 1546716 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 Q8CFP6:p.Ser30Asn mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) n/a 1546717 competitor competitor competitor prey prey chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1546715 unspecified role unspecified role prey prey region 32p radiolabel 32p radiolabel P32 32P 32P P32 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false chemical synthesis Chemical synthesis (Chemical synthesis) direct interaction direct interaction false false false Supp. 2C dnajc27-gtp-5 1546713 n/a 1546715 unspecified role unspecified role prey prey region 32p radiolabel 32p radiolabel P32 32P 32P P32 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1546716 unspecified role unspecified role bait bait Q8CFP6:p.Ser30Asn mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false small_gtp_binding_domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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) direct interaction direct interaction false false false Supp 2C dnajc27-gtp-5 1546713 n/a 1546716 unspecified role unspecified role bait bait Q8CFP6:p.Ser30Asn mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false 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 small_gtp_binding_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 1546715 unspecified role unspecified role prey prey region 32p radiolabel 32p radiolabel P32 32P 32P P32 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1546715 competitor competitor competitor prey prey chemical synthesis Chemical synthesis (Chemical synthesis) direct interaction direct interaction false false false Supp 2C dnajc27-gtp-2 1546714 n/a 1546716 enzyme enzyme neutral component neutral component Q8CFP6:p.His75Leu mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain false Q8CFP6:p.His75Gln mutation increasing rate mutation increasing interaction rate mutation increasing rate certain certain sequence position certain certain certain sequence position certain false 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 1546715 enzyme target enzyme target substrate substrate neutral component neutral component region 32p radiolabel 32p radiolabel P32 32P 32P P32 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false chemical synthesis Chemical synthesis (Chemical synthesis) gtpase reaction gtpase reaction false false false 2B, Supp. 2D dnajc27-gtp-6 1546714 n/a 1546721 enzyme enzyme neutral component neutral component 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 1546715 enzyme target enzyme target substrate substrate neutral component neutral component region 32p radiolabel 32p radiolabel P32 32P 32P P32 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false chemical synthesis Chemical synthesis (Chemical synthesis) gtpase reaction gtpase reaction false false false Supp. 2D