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/ Nm23-H1 metastasis suppressor expression level influences the binding properties, stability, and function of the kinase suppressor of Ras1 (KSR1) Erk scaffold in breast carcinoma cells. Nm23-H1 metastasis suppressor expression level influences the binding properties, stability, and function of the kinase suppressor of Ras1 (KSR1) Erk scaffold in breast carcinoma cells. Mol. Cell. Biol. (0270-7306) 2005 imex curation Salerno M., Palmieri D., Bouadis A., Halverson D., Steeg PS. NDPK - Interactions involving proteins containing InterPro domain IPR001564, Nucleoside diphosphate kinase, core. Cancer - Interactions investigated in the context of cancer The coimmunoprecipitation was carried out with lysates from MDA-MB-435 control cells stably expressing a vector construct (C-100) and cells expressing wild-type nm23-H1 (H1-177). Unique UniProtKB entries could not be associated with Hsp90 and MEK1/2 since the exact gene product of the interactor cannot be ascertained as the antibodies used recognized more than 1 protein. maxsal@mail.nih.gov 13-Nov-2007: Contacted by JYOTI. davet Only protein-protein interactions human-mda_mb_435 Human cell line derived from the M14 melanoma cell line mda_mb_435 Human cell line derived from the M14 melanoma cell line This cell line is now thought to be derived from the M14 melanoma cell line. This cell line was originally thought to be a Human epithelial breast cancer cell line. see pmid: 17004106 anti bait coip anti bait coimmunoprecipitation anti bait coip anti tag western anti tag western blot anti tag western Salerno M., Palmieri D., Bouadis A., Halverson D., Steeg PS. NDPK - Interactions involving proteins containing InterPro domain IPR001564, Nucleoside diphosphate kinase, core. The coimmunoprecipitation was carried out with lysates from MDA-MB-435 control cells stably expressing a vector construct (C-100) and cells expressing wild-type nm23-H1 (H1-177). Unique UniProtKB entries could not be associated with Hsp90 and MEK1/2 since the exact gene product of the interactor cannot be ascertained as the antibodies used recognized more than 1 protein. maxsal@mail.nih.gov Accepted 2007-NOV-3 by DAVET. Mol. Cell. Biol. (0270-7306) 2005 davet Only protein-protein interactions imex curation Nm23-H1 metastasis suppressor expression level influences the binding properties, stability, and function of the kinase suppressor of Ras1 (KSR1) Erk scaffold in breast carcinoma cells. Nm23-H1 metastasis suppressor expression level influences the binding properties, stability, and function of the kinase suppressor of Ras1 (KSR1) Erk scaffold in breast carcinoma cells. Mol. Cell. Biol. (0270-7306) 2005 imex curation Salerno M., Palmieri D., Bouadis A., Halverson D., Steeg PS. NDPK - Interactions involving proteins containing InterPro domain IPR001564, Nucleoside diphosphate kinase, core. Cancer - Interactions investigated in the context of cancer The coimmunoprecipitation was carried out with lysates from MDA-MB-435 control cells stably expressing a vector construct (C-100) and cells expressing wild-type nm23-H1 (H1-177). Unique UniProtKB entries could not be associated with Hsp90 and MEK1/2 since the exact gene product of the interactor cannot be ascertained as the antibodies used recognized more than 1 protein. maxsal@mail.nih.gov 13-Nov-2007: Contacted by JYOTI. davet Only protein-protein interactions human-mda_mb_435 Human cell line derived from the M14 melanoma cell line mda_mb_435 Human cell line derived from the M14 melanoma cell line This cell line is now thought to be derived from the M14 melanoma cell line. This cell line was originally thought to be a Human epithelial breast cancer cell line. see pmid: 17004106 anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Salerno M., Palmieri D., Bouadis A., Halverson D., Steeg PS. NDPK - Interactions involving proteins containing InterPro domain IPR001564, Nucleoside diphosphate kinase, core. The coimmunoprecipitation was carried out with lysates from MDA-MB-435 control cells stably expressing a vector construct (C-100) and cells expressing wild-type nm23-H1 (H1-177). Unique UniProtKB entries could not be associated with Hsp90 and MEK1/2 since the exact gene product of the interactor cannot be ascertained as the antibodies used recognized more than 1 protein. maxsal@mail.nih.gov Accepted 2007-NOV-3 by DAVET. 13-Nov-2007: Contacted by JYOTI. Mol. Cell. Biol. (0270-7306) 2005 davet Only protein-protein interactions imex curation ksr1_mouse Kinase suppressor of Ras 1 Ksr1 Ksr Protein Hb protein protein mouse Mus musculus Mouse MDRAALRAAAMGEKKEGGGGGAAADGGAGAAVSRALQQCGQLQKLIDISIGSLRGLRTKCSVSNDLTQQEIRTLEAKLVKYICKQQQSKLSVTPSDRTAELNSYPRFSDWLYIFNVRPEVVQEIPQELTLDALLEMDEAKAKEMLRRWGASTEECSRLQQALTCLRKVTGLGGEHKMDSGWSSTDARDSSLGPPMDMLSSLGRAGASTQGPRSISVSALPASDSPVPGLSEGLSDSCIPLHTSGRLTPRALHSFITPPTTPQLRRHAKLKPPRTPPPPSRKVFQLLPSFPTLTRSKSHESQLGNRIDDVTPMKFELPHGSPQLVRRDIGLSVTHRFSTKSWLSQVCNVCQKSMIFGVKCKHCRLKCHNKCTKEAPACRITFLPLARLRRTESVPSDINNPVDRAAEPHFGTLPKALTKKEHPPAMNLDSSSNPSSTTSSTPSSPAPFLTSSNPSSATTPPNPSPGQRDSRFSFPDISACSQAAPLSSTADSTRLDDQPKTDVLGVHEAEAEEPEAGKSEAEDDEEDEVDDLPSSRRPWRGPISRKASQTSVYLQEWDIPFEQVELGEPIGQGRWGRVHRGRWHGEVAIRLLEMDGHNQDHLKLFKKEVMNYRQTRHENVVLFMGACMNPPHLAIITSFCKGRTLHSFVRDPKTSLDINKTRQIAQEIIKGMGYLHAKGIVHKDLKSKNVFYDNGKVVITDFGLFGISGVVREERRENQLKLSHDWLCYLAPEIVREMIPGRDEDQLPFSKAADVYAFGTVWYELQARDWPFKHQPAEALIWQIGSGEGVRRVLASVSLGKEVGEILSACWAFDLQERPSFSLLMDMLERLPKLNRRLSHPGHFWKSADINSSKVMPRFERFGLGTLESGNPKM EAEEB23FAE715D94 ndka_human Nucleoside diphosphate kinase A Tumor metastatic process-associated protein Metastasis inhibition factor nm23 Granzyme A-activated DNase NME1 NDPKA NM23 NM23-H1 protein protein human Homo sapiens Human MANCERTFIAIKPDGVQRGLVGEIIKRFEQKGFRLVGLKFMQASEDLLKEHYVDLKDRPFFAGLVKYMHSGPVVAMVWEGLNVVKTGRVMLGETNPADSKPGTIRGDFCIQVGRNIIHGSDSVESAEKEIGLWFHPEELVDYTSCAQNWIYE AAE9C0DF63CB70A1 1433b_human 14-3-3 protein beta/alpha YWHAB Protein kinase C inhibitor protein 1 Protein 1054 protein protein human Homo sapiens Human MTMDKSELVQKAKLAEQAERYDDMAAAMKAVTEQGHELSNEERNLLSVAYKNVVGARRSSWRVISSIEQKTERNEKKQQMGKEYREKIEAELQDICNDVLELLDKYLIPNATQPESKVFYLKMKGDYFRYLSEVASGDNKQTTVSNSQQAYQEAFEISKKEMQPTHPIRLGLALNFSVFYYEILNSPEKACSLAKTAFDEAIAELDTLNEESYKDSTLIMQLLRDNLTLWTSENQGDEGDAGEGEN DIP protein P31946 original sequence version: 171 DIP protein P31946 original sequence version: 97 DIP protein P31946 original sequence version: 119 DIP protein P31946 original sequence version: 122 DIP protein P31946 original sequence version: 180 6BE1A9BF97468017 nme1-ksr1 1 4 unspecified role unspecified role bait bait 3 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 physical association physical association 1 C The data also demonstrate three-way complex formation between Nm23-H1, KSR1, and Hsp90 with either anti-HA-KSR1 or anti-Nm23 as immunoprecipitating antibodies Hsp90 could not be added as an interactor as the antibody used recognized more than 1 isoform of this protein. Q61097; P15531 transfection The interaction occurred in lysates from cells expressing a vector contruct (C-100) or wild-type nm23-H1 (H1-177). P15531 overexpression ksr1-nme1 2 5 unspecified role unspecified role prey prey 3 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 4 unspecified role unspecified role prey prey association association 2 A In the experiment shown in Fig. 2A, the cells were transiently transfected with HAKSR1, and KSR1 was immunoprecipitated from cytoplasmic lysates. Western blot analysis of the KSR1 immunoprecipitations was performed to ascertain relative binding levels of other KSR1 scaffold proteins. Increased binding of Hsp90 to KSR1 was again observed with the H1-177 cell line. In addition, a low level of increased Mek1-2 binding but equivalent 14-3-3 binding were observed with the H1-177 cells. Hsp90 and MEK1/2 could not be added as interactors as the antibody used recognized more than 1 protein. Q61097; P15531 transfection The interaction occurred in lysates from cells expressing a vector contruct (C-100) or wild-type nm23-H1 (H1-177). ksr1-nme1-1 2 4 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 3 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association 3 B Hsp90 could not be added as an interactor as the antibody used recognized more than 1 isoform of this protein. In the experiment shown in Fig. 3B, comparable amounts of KSR1 were immunoprecipitated from cytoplasmic lysates of Nm23-H1-Flag/HA-KSR1- and control Flag/HA-KSR1-transfected cells. Coimmunoprecipitation of KSR1 and endogenous and Nm23-H1-Flag was observed and was more pronounced in the Nm23-H1-Flag/HA-KSR1 transfectants. Q61097; P15531 transfection Endogenous Nm23-H1 was also coimmunoprecipitated with HA-KSR1. Coimmunoprecipitation of KSR1 and endogenous and Nm23-H1-Flag was more pronounced in the Nm23-H1-Flag/HA-KSR1 transfectants. ksr1-nme1-2 2 3 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 4 unspecified role unspecified role prey prey physical association physical association 3 B Hsp90 could not be added as an interactor as the antibody used recognized more than 1 isoform of this protein. In the experiment shown in Fig. 3B, comparable amounts of KSR1 were immunoprecipitated from cytoplasmic lysates of Nm23-H1-Flag/HA-KSR1- and control Flag/HA-KSR1-transfected cells. Coimmunoprecipitation of KSR1 and endogenous and Nm23-H1-Flag was observed and was more pronounced in the Nm23-H1-Flag/HA-KSR1 transfectants. P15531 transfection Coimmunoprecipitation of KSR1 and endogenous and Nm23-H1-Flag was more pronounced in the Nm23-H1-Flag/HA-KSR1 transfectants. ksr1-nme1-3 2 4 unspecified role unspecified role prey prey 3 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association 6 This interaction occured in the presence and absence of 17-AAG, although 17-AAG decreased binding of Hsp90 to KSR1. Hsp90 could not be added as an interactor as the antibody used recognized more than 1 isoform of this protein. This interaction occurred in H1-177 cells (MDA-MB-435 cells stably tranfected with Nm23-H1). It also appears to occur to a lesser extent in control C-100 cells. P15531; Q61097 transfection P15531 overexpression nme1-ksr1-2 1 4 unspecified role unspecified role bait bait 3 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 physical association physical association 1 C The data also demonstrate three-way complex formation between Nm23-H1, KSR1, and Hsp90 with either anti-HA-KSR1 or anti-Nm23 as immunoprecipitating antibodies Hsp90 could not be added as an interactor as the antibody used recognized more than 1 isoform of this protein. Q61097 transfection The interaction occurred in lysates from cells expressing a vector contruct (C-100) or wild-type nm23-H1 (H1-177). ksr1-nme1-4 2 3 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 4 unspecified role unspecified role prey prey physical association physical association 6 This interaction occured in the presence and absence of 17-AAG, although 17-AAG decreased binding of Hsp90 to KSR1. Hsp90 could not be added as an interactor as the antibody used recognized more than 1 isoform of this protein. This interaction occurred in H1-177 cells (MDA-MB-435 cells stably tranfected with Nm23-H1). It also appears to occur to a lesser extent in control C-100 cells. Q61097 transfection