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/ VRK2 inhibits MAPK signaling and inversely correlates with ErbB2 in human breast cancer. VRK2 inhibits MAPK signaling and inversely correlates with ErbB2 in human breast cancer. Mol. Cell. Biol. (0270-7306) 2010 imex curation Fernandez IF., Blanco S., Lozano J., Lazo PA. Cancer - Interactions investigated in the context of cancer 2010-8-17: Lazo PA, Centro de Investigacion del Cancer, CSIC- Universidad de Salamanca, Campus Miguel de Unamuno, E-37007 Salamanca, Spain. plazozbi@usal.es Only protein-protein interactions human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 pull down pull down western blot western blot Immuno blot Immuno blot Fernandez IF., Blanco S., Lozano J., Lazo PA. Cancer - Interactions investigated in the context of cancer 2010-8-17: Lazo PA, Centro de Investigacion del Cancer, CSIC- Universidad de Salamanca, Campus Miguel de Unamuno, E-37007 Salamanca, Spain. check the supplemental material when available plazozbi@usal.es Accepted 2010-SEP-18 by MGT. Mol. Cell. Biol. (0270-7306) 2010 Only protein-protein interactions imex curation VRK2 inhibits MAPK signaling and inversely correlates with ErbB2 in human breast cancer. VRK2 inhibits MAPK signaling and inversely correlates with ErbB2 in human breast cancer. Mol. Cell. Biol. (0270-7306) 2010 imex curation Fernandez IF., Blanco S., Lozano J., Lazo PA. Cancer - Interactions investigated in the context of cancer 2010-8-17: Lazo PA, Centro de Investigacion del Cancer, CSIC- Universidad de Salamanca, Campus Miguel de Unamuno, E-37007 Salamanca, Spain. plazozbi@usal.es 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 western blot western blot Immuno blot Immuno blot Fernandez IF., Blanco S., Lozano J., Lazo PA. Cancer - Interactions investigated in the context of cancer 2010-8-17: Lazo PA, Centro de Investigacion del Cancer, CSIC- Universidad de Salamanca, Campus Miguel de Unamuno, E-37007 Salamanca, Spain. plazozbi@usal.es Accepted 2010-SEP-18 by MGT. Mol. Cell. Biol. (0270-7306) 2010 Only protein-protein interactions imex curation VRK2 inhibits MAPK signaling and inversely correlates with ErbB2 in human breast cancer. VRK2 inhibits MAPK signaling and inversely correlates with ErbB2 in human breast cancer. Mol. Cell. Biol. (0270-7306) 2010 imex curation Fernandez IF., Blanco S., Lozano J., Lazo PA. Cancer - Interactions investigated in the context of cancer 2010-8-17: Lazo PA, Centro de Investigacion del Cancer, CSIC- Universidad de Salamanca, Campus Miguel de Unamuno, E-37007 Salamanca, Spain. plazozbi@usal.es Only protein-protein interactions human-mcf7 Homo sapiens mammary carcinoma cells mcf7 mammary carcinoma cells coip coimmunoprecipitation co-immunoprecipitation CoIp immunoprecipitation Co-IP co-immunoprecipitation CoIp immunoprecipitation Co-IP coip western blot western blot Immuno blot Immuno blot Fernandez IF., Blanco S., Lozano J., Lazo PA. Cancer - Interactions investigated in the context of cancer 2010-8-17: Lazo PA, Centro de Investigacion del Cancer, CSIC- Universidad de Salamanca, Campus Miguel de Unamuno, E-37007 Salamanca, Spain. plazozbi@usal.es Accepted 2010-SEP-18 by MGT. Mol. Cell. Biol. (0270-7306) 2010 Only protein-protein interactions imex curation VRK2 inhibits MAPK signaling and inversely correlates with ErbB2 in human breast cancer. VRK2 inhibits MAPK signaling and inversely correlates with ErbB2 in human breast cancer. Mol. Cell. Biol. (0270-7306) 2010 imex curation Fernandez IF., Blanco S., Lozano J., Lazo PA. Cancer - Interactions investigated in the context of cancer 2010-8-17: Lazo PA, Centro de Investigacion del Cancer, CSIC- Universidad de Salamanca, Campus Miguel de Unamuno, E-37007 Salamanca, Spain. plazozbi@usal.es Only protein-protein interactions human-mda_mb_231 Homo sapiens Caucasian breast adenocarcinoma mda_mb_231 Human Caucasian breast adenocarcinoma confocal microscopy confocal microscopy antibody detection identification by antibody antibody detection Fernandez IF., Blanco S., Lozano J., Lazo PA. Cancer - Interactions investigated in the context of cancer 2010-8-17: Lazo PA, Centro de Investigacion del Cancer, CSIC- Universidad de Salamanca, Campus Miguel de Unamuno, E-37007 Salamanca, Spain. plazozbi@usal.es Accepted 2010-SEP-18 by MGT. Mol. Cell. Biol. (0270-7306) 2010 Only protein-protein interactions imex curation VRK2 inhibits MAPK signaling and inversely correlates with ErbB2 in human breast cancer. VRK2 inhibits MAPK signaling and inversely correlates with ErbB2 in human breast cancer. Mol. Cell. Biol. (0270-7306) 2010 imex curation Fernandez IF., Blanco S., Lozano J., Lazo PA. Cancer - Interactions investigated in the context of cancer 2010-8-17: Lazo PA, Centro de Investigacion del Cancer, CSIC- Universidad de Salamanca, Campus Miguel de Unamuno, E-37007 Salamanca, Spain. plazozbi@usal.es Only protein-protein interactions human-mcf7 Homo sapiens mammary carcinoma cells mcf7 mammary carcinoma cells confocal microscopy confocal microscopy antibody detection identification by antibody antibody detection Fernandez IF., Blanco S., Lozano J., Lazo PA. Cancer - Interactions investigated in the context of cancer 2010-8-17: Lazo PA, Centro de Investigacion del Cancer, CSIC- Universidad de Salamanca, Campus Miguel de Unamuno, E-37007 Salamanca, Spain. plazozbi@usal.es Accepted 2010-SEP-18 by MGT. Mol. Cell. Biol. (0270-7306) 2010 Only protein-protein interactions imex curation VRK2 inhibits MAPK signaling and inversely correlates with ErbB2 in human breast cancer. VRK2 inhibits MAPK signaling and inversely correlates with ErbB2 in human breast cancer. Mol. Cell. Biol. (0270-7306) 2010 imex curation Fernandez IF., Blanco S., Lozano J., Lazo PA. Cancer - Interactions investigated in the context of cancer 2010-8-17: Lazo PA, Centro de Investigacion del Cancer, CSIC- Universidad de Salamanca, Campus Miguel de Unamuno, E-37007 Salamanca, Spain. plazozbi@usal.es Only protein-protein interactions human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 confocal microscopy confocal microscopy antibody detection identification by antibody antibody detection Fernandez IF., Blanco S., Lozano J., Lazo PA. Cancer - Interactions investigated in the context of cancer 2010-8-17: Lazo PA, Centro de Investigacion del Cancer, CSIC- Universidad de Salamanca, Campus Miguel de Unamuno, E-37007 Salamanca, Spain. plazozbi@usal.es Accepted 2010-SEP-18 by MGT. Mol. Cell. Biol. (0270-7306) 2010 Only protein-protein interactions imex curation VRK2 inhibits MAPK signaling and inversely correlates with ErbB2 in human breast cancer. VRK2 inhibits MAPK signaling and inversely correlates with ErbB2 in human breast cancer. Mol. Cell. Biol. (0270-7306) 2010 imex curation Fernandez IF., Blanco S., Lozano J., Lazo PA. Cancer - Interactions investigated in the context of cancer 2010-8-17: Lazo PA, Centro de Investigacion del Cancer, CSIC- Universidad de Salamanca, Campus Miguel de Unamuno, E-37007 Salamanca, Spain. plazozbi@usal.es 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 western blot western blot Immuno blot Immuno blot Fernandez IF., Blanco S., Lozano J., Lazo PA. Cancer - Interactions investigated in the context of cancer 2010-8-17: Lazo PA, Centro de Investigacion del Cancer, CSIC- Universidad de Salamanca, Campus Miguel de Unamuno, E-37007 Salamanca, Spain. plazozbi@usal.es Accepted 2010-SEP-18 by MGT. Mol. Cell. Biol. (0270-7306) 2010 Only protein-protein interactions imex curation calx_human Calnexin CANX Major histocompatibility complex class I antigen-binding protein p88 p90 IP90 protein protein human Homo sapiens Human MEGKWLLCMLLVLGTAIVEAHDGHDDDVIDIEDDLDDVIEEVEDSKPDTTAPPSSPKVTYKAPVPTGEVYFADSFDRGTLSGWILSKAKKDDTDDEIAKYDGKWEVEEMKESKLPGDKGLVLMSRAKHHAISAKLNKPFLFDTKPLIVQYEVNFQNGIECGGAYVKLLSKTPELNLDQFHDKTPYTIMFGPDKCGEDYKLHFIFRHKNPKTGIYEEKHAKRPDADLKTYFTDKKTHLYTLILNPDNSFEILVDQSVVNSGNLLNDMTPPVNPSREIEDPEDRKPEDWDERPKIPDPEAVKPDDWDEDAPAKIPDEEATKPEGWLDDEPEYVPDPDAEKPEDWDEDMDGEWEAPQIANPRCESAPGCGVWQRPVIDNPNYKGKWKPPMIDNPSYQGIWKPRKIPNPDFFEDLEPFRMTPFSAIGLELWSMTSDIFFDNFIICADRRIVDDWANDGWGLKKAADGAAEPGVVGQMIEAAEERPWLWVVYILTVALPVFLVILFCCSGKKQTSGMEYKKTDAPQPDVKEEEEEKEEEKDKGDEEEEGEEKLEEKQKSDAEEDGGTVSQEEEDRKPKAEEDEILNRSPRNRKPRRE DIP protein P27824 original sequence version: 177 EDE094D9B82261EE vrk2_human Serine/threonine-protein kinase VRK2 VRK2 Vaccinia-related kinase 2 protein protein human Homo sapiens Human MPPKRNEKYKLPIPFPEGKVLDDMEGNQWVLGKKIGSGGFGLIYLAFPTNKPEKDARHVVKVEYQENGPLFSELKFYQRVAKKDCIKKWIERKQLDYLGIPLFYGSGLTEFKGRSYRFMVMERLGIDLQKISGQNGTFKKSTVLQLGIRMLDVLEYIHENEYVHGDIKAANLLLGYKNPDQVYLADYGLSYRYCPNGNHKQYQENPRKGHNGTIEFTSLDAHKGVALSRRSDVEILGYCMLRWLCGKLPWEQNLKDPVAVQTAKTNLLDELPQSVLKWAPSGSSCCEIAQFLVCAHSLAYDEKPNYQALKKILNPHGIPLGPLDFSTKGQSINVHTPNSQKVDSQKAATKQVNKAHNRLIEKKVHSERSAESCATWKVQKEEKLIGLMNNEAAQESTRRRQKYQESQEPLNEVNSFPQKISYTQFPNSFYEPHQDFTSPDIFKKSRSPSWYKYTSTVSTGITDLESSTGLWPTISQFTLSEETNADVYYYRIIIPVLLMLVFLALFFL 9F3F6FCC9280568F mp2k1_human Dual specificity mitogen-activated protein kinase kinase 1 ERK activator kinase 1 MAPK/ERK kinase 1 MAP2K1 MEK1 PRKMK1 protein protein human Homo sapiens Human MPKKKPTPIQLNPAPDGSAVNGTSSAETNLEALQKKLEELELDEQQRKRLEAFLTQKQKVGELKDDDFEKISELGAGNGGVVFKVSHKPSGLVMARKLIHLEIKPAIRNQIIRELQVLHECNSPYIVGFYGAFYSDGEISICMEHMDGGSLDQVLKKAGRIPEQILGKVSIAVIKGLTYLREKHKIMHRDVKPSNILVNSRGEIKLCDFGVSGQLIDSMANSFVGTRSYMSPERLQGTHYSVQSDIWSMGLSLVEMAVGRYPIPPPDAKELELMFGCQVEGDAAETPPRPRTPGRPLSSYGMDSRPPMAIFELLDYIVNEPPPKLPSGVFSLEFQDFVNKCLIKNPAERADLKQLMVHAFIKRSDAEEVDFAGWLCSTIGLNQPSTPTHAAGV DIP protein Q02750 original sequence version: 96 DIP protein Q02750 original sequence version: 93 DIP protein Q02750-1 original sequence version: 151 DIP protein Q02750 original sequence version: 151 DIP protein Q02750-1 original sequence version: 166 0344118FFC842D51 ksr1_human Kinase suppressor of Ras 1 KSR1 KSR protein protein human Homo sapiens Human MDRAALRAAAMGEKKEGGGGGDAAAAEGGAGAAASRALQQCGQLQKLIDISIGSLRGLRTKCAVSNDLTQQEIRTLEAKLVRYICKQRQCKLSVAPGERTPELNSYPRFSDWLYTFNVRPEVVQEIPRDLTLDALLEMNEAKVKETLRRCGASGDECGRLQYALTCLRKVTGLGGEHKEDSSWSSLDARRESGSGPSTDTLSAASLPWPPGSSQLGRAGNSAQGPRSISVSALPASDSPTPSFSEGLSDTCIPLHASGRLTPRALHSFITPPTTPQLRRHTKLKPPRTPPPPSRKVFQLLPSFPTLTRSKSHESQLGNRIDDVSSMRFDLSHGSPQMVRRDIGLSVTHRFSTKSWLSQVCHVCQKSMIFGVKCKHCRLKCHNKCTKEAPACRISFLPLTRLRRTESVPSDINNPVDRAAEPHFGTLPKALTKKEHPPAMNHLDSSSNPSSTTSSTPSSPAPFPTSSNPSSATTPPNPSPGQRDSRFNFPAAYFIHHRQQFIFPVPSAGHCWKCLLIAESLKENAFNISAFAHAAPLPEAADGTRLDDQPKADVLEAHEAEAEEPEAGKSEAEDDEDEVDDLPSSRRPWRGPISRKASQTSVYLQEWDIPFEQVELGEPIGQGRWGRVHRGRWHGEVAIRLLEMDGHNQDHLKLFKKEVMNYRQTRHENVVLFMGACMNPPHLAIITSFCKGRTLHSFVRDPKTSLDINKTRQIAQEIIKGMGYLHAKGIVHKDLKSKNVFYDNGKVVITDFGLFGISGVVREGRRENQLKLSHDWLCYLAPEIVREMTPGKDEDQLPFSKAADVYAFGTVWYELQARDWPLKNQAAEASIWQIGSGEGMKRVLTSVSLGKEVSEILSACWAFDLQERPSFSLLMDMLEKLPKLNRRLSHPGHFWKSADINSSKVVPRFERFGLGVLESSNPKM DIP protein Q8IVT5 original sequence version: 115 61BF0F1598349540 ksr1-calnexin 6 8 unspecified role unspecified role neutral component neutral component 11 unspecified role unspecified role neutral component neutral component colocalization colocalization 6A The subcelular location of KSR1 in the cytoplasm is partly dispersed and partly colocalizing with endogenous calnexin, an endoplasmic reticulum marker vrk2-ksr1 6 11 unspecified role unspecified role neutral component neutral component 9 unspecified role unspecified role neutral component neutral component colocalization colocalization 6B This VRK2-KSR1 colocalization was confirmed by immunofluorescence (Fig 6B). ksr1-calnexin-1 5 8 unspecified role unspecified role neutral component neutral component 11 unspecified role unspecified role neutral component neutral component colocalization colocalization 6A The subcelular location of KSR1 in the cytoplasm is partly dispersed and partly colocalizing with endogenous calnexin, an endoplasmic reticulum marker ksr1-calnexin-2 4 11 unspecified role unspecified role neutral component neutral component 8 unspecified role unspecified role neutral component neutral component colocalization colocalization 6A The subcelular location of KSR1 in the cytoplasm is partly dispersed and partly colocalizing with endogenous calnexin, an endoplasmic reticulum marker vrk2a-ksr1 7 11 unspecified role unspecified role prey prey 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 7A First a direct interaction KSR1-VRK2A interaction could be detected by immunoprecipitation of transfected KSR1, which was clearly bound to VRK2A (Fig. 7A left), and also in a reciprocal experiment by a pulldown of VRK2 which binds to endogenous KSR1 (Fig. 7A right). ksr1-vrk2a 2 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 11 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 7A First a direct interaction KSR1-VRK2A interaction could be detected by immunoprecipitation of transfected KSR1, which was clearly bound to VRK2A (Fig. 7A left), and also in a reciprocal experiment by a pulldown of VRK2 which binds to endogenous KSR1 (Fig. 7A right). vrk2a-mek 1 10 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 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 7B Only MEK stably interacted with VRK2A (Fig. 7B) mek1-vrk2a 3 10 unspecified role unspecified role bait bait 9 unspecified role unspecified role prey prey physical association physical association 7C The interaction between endogenous VRK2 and MEK1 was also determined and detected by immunoprecipitation (Fig. 7C)