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/ Apoptosis signal-regulating kinase (ASK) 2 functions as a mitogen-activated protein kinase kinase kinase in a heteromeric complex with ASK1. Apoptosis signal-regulating kinase (ASK) 2 functions as a mitogen-activated protein kinase kinase kinase in a heteromeric complex with ASK1. J. Biol. Chem. (0021-9258) 2007 imex curation Takeda K., Shimozono R., Noguchi T., Umeda T., Morimoto Y., Naguro I., Tobiume K., Saitoh M., Matsuzawa A., Ichijo H. Apoptosis - Interactions involving proteins with a function related to apoptosis Species of origin of MAP3K5 was determined by mutant mapping of construct described in paper and in PMID: 9564042 ichijo@mol.f.u-tokyo.ac.jp 29-OCT-2007: Contacted by JYOTI. Human fibroblast WI-38cDNA Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Takeda K., Shimozono R., Noguchi T., Umeda T., Morimoto Y., Naguro I., Tobiume K., Saitoh M., Matsuzawa A., Ichijo H. Apoptosis - Interactions involving proteins with a function related to apoptosis Species of origin of MAP3K5 was determined by mutant mapping of construct described in paper and in PMID: 9564042 ichijo@mol.f.u-tokyo.ac.jp Accepted 2007-JUL-13 by JYOTI. Human fibroblast WI-38cDNA J. Biol. Chem. (0021-9258) 2007 Only protein-protein interactions imex curation Apoptosis signal-regulating kinase (ASK) 2 functions as a mitogen-activated protein kinase kinase kinase in a heteromeric complex with ASK1. Apoptosis signal-regulating kinase (ASK) 2 functions as a mitogen-activated protein kinase kinase kinase in a heteromeric complex with ASK1. J. Biol. Chem. (0021-9258) 2007 imex curation Takeda K., Shimozono R., Noguchi T., Umeda T., Morimoto Y., Naguro I., Tobiume K., Saitoh M., Matsuzawa A., Ichijo H. Apoptosis - Interactions involving proteins with a function related to apoptosis Species of origin of MAP3K5 was determined by mutant mapping of construct described in paper and in PMID: 9564042 ichijo@mol.f.u-tokyo.ac.jp 29-OCT-2007: Contacted by JYOTI. Human fibroblast WI-38cDNA Only protein-protein interactions 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 Takeda K., Shimozono R., Noguchi T., Umeda T., Morimoto Y., Naguro I., Tobiume K., Saitoh M., Matsuzawa A., Ichijo H. Apoptosis - Interactions involving proteins with a function related to apoptosis Species of origin of MAP3K5 was determined by mutant mapping of construct described in paper and in PMID: 9564042 ichijo@mol.f.u-tokyo.ac.jp Accepted 2007-JUL-13 by JYOTI. 29-OCT-2007: Contacted by JYOTI. Human fibroblast WI-38cDNA J. Biol. Chem. (0021-9258) 2007 Only protein-protein interactions imex curation Apoptosis signal-regulating kinase (ASK) 2 functions as a mitogen-activated protein kinase kinase kinase in a heteromeric complex with ASK1. Apoptosis signal-regulating kinase (ASK) 2 functions as a mitogen-activated protein kinase kinase kinase in a heteromeric complex with ASK1. J. Biol. Chem. (0021-9258) 2007 imex curation Takeda K., Shimozono R., Noguchi T., Umeda T., Morimoto Y., Naguro I., Tobiume K., Saitoh M., Matsuzawa A., Ichijo H. Apoptosis - Interactions involving proteins with a function related to apoptosis Species of origin of MAP3K5 was determined by mutant mapping of construct described in paper and in PMID: 9564042 ichijo@mol.f.u-tokyo.ac.jp 29-OCT-2007: Contacted by JYOTI. Human fibroblast WI-38cDNA Only protein-protein interactions mouse-embryonic_fibroblast Mus musculus embryonic fibroblasts embryonic_fibroblast Embryonic fibroblast Fibroblasts taken from an embryonic as opposed to a foetal or adult source. Foetus is defined as the stage at which tissue diferentiation is initiated. Tissue taken from an embryonic as opposed to a foetal or adult source. Embryo is defined as the development stage prior to organ formation. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Takeda K., Shimozono R., Noguchi T., Umeda T., Morimoto Y., Naguro I., Tobiume K., Saitoh M., Matsuzawa A., Ichijo H. Apoptosis - Interactions involving proteins with a function related to apoptosis Species of origin of MAP3K5 was determined by mutant mapping of construct described in paper and in PMID: 9564042 ichijo@mol.f.u-tokyo.ac.jp Accepted 2007-JUL-13 by JYOTI. 29-OCT-2007: Contacted by JYOTI. Human fibroblast WI-38cDNA J. Biol. Chem. (0021-9258) 2007 Only protein-protein interactions imex curation Apoptosis signal-regulating kinase (ASK) 2 functions as a mitogen-activated protein kinase kinase kinase in a heteromeric complex with ASK1. Apoptosis signal-regulating kinase (ASK) 2 functions as a mitogen-activated protein kinase kinase kinase in a heteromeric complex with ASK1. J. Biol. Chem. (0021-9258) 2007 imex curation Takeda K., Shimozono R., Noguchi T., Umeda T., Morimoto Y., Naguro I., Tobiume K., Saitoh M., Matsuzawa A., Ichijo H. Apoptosis - Interactions involving proteins with a function related to apoptosis Species of origin of MAP3K5 was determined by mutant mapping of construct described in paper and in PMID: 9564042 ichijo@mol.f.u-tokyo.ac.jp 29-OCT-2007: Contacted by JYOTI. Human fibroblast WI-38cDNA Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid predetermined predetermined participant predetermined Takeda K., Shimozono R., Noguchi T., Umeda T., Morimoto Y., Naguro I., Tobiume K., Saitoh M., Matsuzawa A., Ichijo H. Apoptosis - Interactions involving proteins with a function related to apoptosis Species of origin of MAP3K5 was determined by mutant mapping of construct described in paper and in PMID: 9564042 ichijo@mol.f.u-tokyo.ac.jp Accepted 2007-JUL-13 by JYOTI. 29-OCT-2007: Contacted by JYOTI. Human fibroblast WI-38cDNA J. Biol. Chem. (0021-9258) 2007 Only protein-protein interactions imex curation m3k6_mouse Mitogen-activated protein kinase kinase kinase 6 Map3k6 Ask2 Mapkkk6 Mekk6 Apoptosis signal-regulating kinase 2 protein protein mouse Mus musculus Mouse MAGPCPRAGVLERAGSCWQDPLAEALSRGRSSPAVTGRGCARSRPLSVVYVLTREPGPGVEPGSGTEAEPLPLRCLREACAQLQGTRPPPQLRSLPFATLALGDTAALDSFYNADVVVLEVSSSLAQPSLFYHLGVRESFSMTNNVLLCSQAELPDLQALREDVFQKNSDCVGSYTLIPYVVTATGRVLCGDAGLLRGIADGLVQAGAGTEALLTPLVGRLVRLLEATPTDSCGYFRETIRQDIRQARERFSGQQLRQELARLQRRLDSVELLSPDIVMNLLLSYRDVQDYSAIIELVETLQALPTCDVAEQHNVCFHYTFALNRRNRPGDREKALAVLLPLVKYEGPVAPDLYCMCGRIYKDMFFTSGFQNAGHLEQAYHWYRKAFDVEPSLHSGINAAVLLIAAGQHFEDSEELRLIGMKLACLLARKGCVEKMQYYWDVGFYLGAQILANDPIQVVLAAEQLYKLNAPIWYLVSVMETFLLYQHFRPTPEPSGGPPLRAHFWLHFLLQSCQPFKMAPPQEDQCLVLVLEINKVLLPARLEIQGTDPMSAVTLSLLEPETQEDPSSWTFPVTSICGISTSKLDQRCCFLYALPPAQDVQLCFPSVERCQRFCGLIQVLVMNPDSSAPTEEAEGAREVLEFDYEYSETGERLVLGRGTYGVVYAGRDRHTRVRIAIKEIPERDSRFSQPLHEEIALHKRLRHKNIVRYLGSASQGGYLKIFMEEVPGGSLSSLLRSVWGPLKDNESTISFYTRQILQGLSYLHENRIVHRDIKGDNVLINTFSGLLKISDFGTSKRLAGITPCTETFTGTLQYMAPEIIDQGPRGYGKAADIWSLGCTVIEMATGRPPFHELGSPQAAMFQVGMYKVHPPVPGSLSAEAQAFLLRTFEPDPRLRASAQELLGDPFLQPGKRSRSPGSPRHTPRPSGAPSGPSSPSADSATQSQTFPRPQAPSQHPPSPPKRCLSYGDTSQLRVPEEPAAEEPASPEESSGLSLLHQESKRRAMLAAVLEQEVPTLAENLLEQEQDSRLSKIHVELLLRCLGAQIHTPNRRQLAQELRTLQAQLRAQSLGPALLKGPLFAFPDAVKQILRRRQIRPHWMFVLDSLLSRAVRAALAVLDAESEKKAVLPRSEESSKEESQQKPQESQALQSQLPPEQGPPSLMVELGLLRAETDRLRDLLAEKERECQALVQQALHRVHAETRKYAPASETPATLPKDQNLVRWLQELSVDPATIQTLLSHSFTLQTLLTCATQDDLVYTRIRGGMVCRIWRAILAQRAGATSVTPVPRDAE FC61A90935A7465D m3k5_human Mitogen-activated protein kinase kinase kinase 5 MAPK/ERK kinase kinase 5 Apoptosis signal-regulating kinase 1 MAP3K5 ASK1 MAPKKK5 MEKK5 protein protein human Homo sapiens Human MSTEADEGITFSVPPFAPSGFCTIPEGGICRRGGAAAVGEGEEHQLPPPPPGSFWNVESAAAPGIGCPAATSSSSATRGRGSSVGGGSRRTTVAYVINEASQGQLVVAESEALQSLREACETVGATLETLHFGKLDFGETTVLDRFYNADIAVVEMSDAFRQPSLFYHLGVRESFSMANNIILYCDTNSDSLQSLKEIICQKNTMCTGNYTFVPYMITPHNKVYCCDSSFMKGLTELMQPNFELLLGPICLPLVDRFIQLLKVAQASSSQYFRESILNDIRKARNLYTGKELAAELARIRQRVDNIEVLTADIVINLLLSYRDIQDYDSIVKLVETLEKLPTFDLASHHHVKFHYAFALNRRNLPGDRAKALDIMIPMVQSEGQVASDMYCLVGRIYKDMFLDSNFTDTESRDHGASWFKKAFESEPTLQSGINYAVLLLAAGHQFESSFELRKVGVKLSSLLGKKGNLEKLQSYWEVGFFLGASVLANDHMRVIQASEKLFKLKTPAWYLKSIVETILIYKHFVKLTTEQPVAKQELVDFWMDFLVEATKTDVTVVRFPVLILEPTKIYQPSYLSINNEVEEKTISIWHVLPDDKKGIHEWNFSASSVRGVSISKFEERCCFLYVLHNSDDFQIYFCTELHCKKFFEMVNTITEEKGRSTEEGDCESDLLEYDYEYDENGDRVVLGKGTYGIVYAGRDLSNQVRIAIKEIPERDSRYSQPLHEEIALHKHLKHKNIVQYLGSFSENGFIKIFMEQVPGGSLSALLRSKWGPLKDNEQTIGFYTKQILEGLKYLHDNQIVHRDIKGDNVLINTYSGVLKISDFGTSKRLAGINPCTETFTGTLQYMAPEIIDKGPRGYGKAADIWSLGCTIIEMATGKPPFYELGEPQAAMFKVGMFKVHPEIPESMSAEAKAFILKCFEPDPDKRACANDLLVDEFLKVSSKKKKTQPKLSALSAGSNEYLRSISLPVPVLVEDTSSSSEYGSVSPDTELKVDPFSFKTRAKSCGERDVKGIRTLFLGIPDENFEDHSAPPSPEEKDSGFFMLRKDSERRATLHRILTEDQDKIVRNLMESLAQGAEEPKLKWEHITTLIASLREFVRSTDRKIIATTLSKLKLELDFDSHGISQVQVVLFGFQDAVNKVLRNHNIKPHWMFALDSIIRKAVQTAITILVPELRPHFSLASESDTADQEDLDVEDDHEEQPSNQTVRRPQAVIEDAVATSGVSTLSSTVSHDSQSAHRSLNVQLGRMKIETNRLLEELVRKEKELQALLHRAIEEKDQEIKHLKLKSQPIEIPELPVFHLNSSGTNTEDSELTDWLRVNGADEDTISRFLAEDYTLLDVLYYVTRDDLKCLRLRGGMLCTLWKAIIDFRNKQT DIP protein Q99683 original sequence version: 83 DIP protein Q99683 original sequence version: 151 DIP protein Q99683 original sequence version: 113 265BDC65968AF985 m3k5_mouse Mitogen-activated protein kinase kinase kinase 5 MAPK/ERK kinase kinase 5 Apoptosis signal-regulating kinase 1 Map3k5 Ask1 Mekk5 protein protein mouse Mus musculus Mouse MGTEAGEGITFSVPPFASVGFCTIPEGGSCRRGGGAATAAEGEPSLQPLLVPPPPPPPGSFWNVESAAAPGTSCPTTAPGSSATRGRGNSGSGGGRRTTVAYVINEASQGQLVVAESEALQSLREACEAVGATLETLHFGKLDFGETAVLDRFYNADIAVVEMSDAFRQPSLFYHLGVRESFSMANNIILYCDTNSDSLQSLKEIICQKNTVCTGNYTFIPYMVTPHNKVYCCDSSFMKGLTELMQPNFELLLGPICLPLVDRFVQLLKVAQASSSQYFRESILSDIRKARNLYTGKELAAELARIRQRVDNIEVLTADIVINLLLSYRDIQDYDSIVKLVETLEKLPTFDLASHHHVKFHYAFALNRRNLPGDRAKALDIMIPMVQSEEQVASDMYCLVGRIYKDMFLDSNFTDTESRDHGASWFKKAFESEPTLQSGINYAVLLLAAGHQFESSFELRKVGVKLSSLLGKKGNLEKLQSYWEVGFFLGASVLANDHLRVIQASEKLFRLKTPAWYLKSIVETILIYKHFVKLTTEQPSAKQELVDFWMDFLVEATKTDVTVVRFPVLILEPTKIYQPSYLSINNEVEEKTISIWHVLPDDKKGIHEWNFGASSVRGVSISKFEERCCFLYVLHNSDDFQIYFCTELHCKRFFEMVNTITEEKGRGAEDGDCEGDSLEYDYEYDENGDRVVLGKGTYGIVYAGRDLSNQVRIAIKEIPERDSRYSQPLHEEIALHKHLKHKNIVQYLGSFSENGFIKIFMEQVPGGSLSALLRSKWGPLKDNEQTIGFYTKQILEGLKYLHDNQIVHRDIKGDNVLINTYSGVLKISDFGTSKRLAGINPCTETFTGTLQYMAPEIIDKGPRGYGKAADIWSLGCTIIEMATGKPPFYELGEPQAAMFKVGMFKVHPEIPESMSAEAKAFILKCFEPDPDKRACANDLLIDEFLKVSSKKKKTQPKLSALSTGSNEYLRSISLPVPVLVEDTSSSSEYGSVSPDTELKADPFSFKARAKSCGEKDGKGIRTLFLGIPDENFEDHSAPPSPEEKDSGFFMLRKDSERRATLHRILTEDQDKVVRNLMESLAQGAEEPKLKWEHITTLISSLREFVRSTDRKIIATTLSKLKLELDFDSHGISQVQVVLFGFQDAVNKVLRNHNIKPHWMFALDSIIRKAVQTAITILVPELRPHFSLASESDTADPEDLDVEDEHEELSSNQTVRRPQAITEDAVATSGVSTLSSTVSHDSQNAHRSLNVQLGRMKIETNRLLEELVRKERELQALLHQAIEEKDQEIRHLKLKSQPIDIPGFPVCHLNSPGTTTEDSELPGWLRENGADEDTISRFLAEDYTLVDVLYYVTRDDLKCLRLRGGMLCTLWKAIIDFRNKC DIP protein O35099 original sequence version: 123 BE68D225EBBCDF38 m3k6_human Mitogen-activated protein kinase kinase kinase 6 MAP3K6 MAPKKK6 ASK2 MEKK6 Apoptosis signal-regulating kinase 2 protein protein human Homo sapiens Human MAGPCPRSGAERAGSCWQDPLAVALSRGRQLAAPPGRGCARSRPLSVVYVLTREPQPGLEPREGTEAEPLPLRCLREACAQVPRPRPPPQLRSLPFGTLELGDTAALDAFYNADVVVLEVSSSLVQPSLFYHLGVRESFSMTNNVLLCSQADLPDLQALREDVFQKNSDCVGSYTLIPYVVTATGRVLCGDAGLLRGLADGLVQAGVGTEALLTPLVGRLARLLEATPTDSCGYFRETIRRDIRQARERFSGPQLRQELARLQRRLDSVELLSPDIIMNLLLSYRDVQDYSAIIELVETLQALPTCDVAEQHNVCFHYTFALNRRNRPGDRAKALSVLLPLVQLEGSVAPDLYCMCGRIYKDMFFSSGFQDAGHREQAYHWYRKAFDVEPSLHSGINAAVLLIAAGQHFEDSKELRLIGMKLGCLLARKGCVEKMQYYWDVGFYLGAQILANDPTQVVLAAEQLYKLNAPIWYLVSVMETFLLYQHFRPTPEPPGGPPRRAHFWLHFLLQSCQPFKTACAQGDQCLVLVLEMNKVLLPAKLEVRGTDPVSTVTLSLLEPETQDIPSSWTFPVASICGVSASKRDERCCFLYALPPAQDVQLCFPSVGHCQWFCGLIQAWVTNPDSTAPAEEAEGAGEMLEFDYEYTETGERLVLGKGTYGVVYAGRDRHTRVRIAIKEIPERDSRFSQPLHEEIALHRRLRHKNIVRYLGSASQGGYLKIFMEEVPGGSLSSLLRSVWGPLKDNESTISFYTRQILQGLGYLHDNHIVHRDIKGDNVLINTFSGLLKISDFGTSKRLAGITPCTETFTGTLQYMAPEIIDQGPRGYGKAADIWSLGCTVIEMATGRPPFHELGSPQAAMFQVGMYKVHPPMPSSLSAEAQAFLLRTFEPDPRLRASAQTLLGDPFLQPGKRSRSPSSPRHAPRPSDAPSASPTPSANSTTQSQTFPCPQAPSQHPPSPPKRCLSYGGTSQLRVPEEPAAEEPASPEESSGLSLLHQESKRRAMLAAVLEQELPALAENLHQEQKQEQGARLGRNHVEELLRCLGAHIHTPNRRQLAQELRALQGRLRAQGLGPALLHRPLFAFPDAVKQILRKRQIRPHWMFVLDSLLSRAVRAALGVLGPEVEKEAVSPRSEELSNEGDSQQSPGQQSPLPVEPEQGPAPLMVQLSLLRAETDRLREILAGKEREYQALVQRALQRLNEEARTYVLAPEPPTALSTDQGLVQWLQELNVDSGTIQMLLNHSFTLHTLLTYATRDDLIYTRIRGGMVCRIWRAILAQRAGSTPVTSGP 013AF3729698080F map3k5-map3k6-1 1 6 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 8 unspecified role unspecified role prey prey physical association physical association To identify molecules that interact with ASK1, we employed the yeast two-hybrid system using a deletion mutant of ASK1 map3k5-map3k6-2 4 5 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait Q99683:p.Lys709Asn mutation mutation certain certain sequence position certain certain certain sequence position certain K N Wed May 02 17:56:35 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys709asn' Tue Jun 30 19:18:07 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys709Asn' physical association physical association The interaction of ASK1 with the cloned full-length version of mouse ASK2 was confirmed by the yeast two-hybrid assay (Fig. 1C). 1C Q99683 mutant is kinase-dead map3k5-map3k6-3 2 5 unspecified role unspecified role bait bait n-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range 6 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association We also detected the interaction between Myc-tagged ASK1 (Myc-ASK1) and FLAG-tagged ASK2 (FLAG-ASK2) by co-immunoprecipitation analysis in HEK293 cells (Fig. 1D). 1D map3k5-map3k6-4 3 5 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait physical association physical association Cell lysates from MEFs were subjected to immunoprecipitation with the ASK1 antibody, and endogenous ASK2 co-purified with ASK1 was clearly detected by immunoblotting with the ASK2 antibody (Fig. 1F). 1F map3k5-map3k6-5 2 5 unspecified role unspecified role bait bait n-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range 6 unspecified role unspecified role prey prey n-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range c-terminal region required to bind necessary binding region required to bind deletion analysis deletion analysis range range certain certain sequence position certain physical association physical association HA-ASK1-CT but not HA-ASK1- was found to be co-precipitated with FLAG-ASK2, indicating that ASK1-CT was necessary and sufficient for the binding of ASK1 to ASK2 (Fig. 3B). 3B