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/ beinke-2003-1 NF-kappaB1 p105 negatively regulates TPL-2 MEK kinase activity. 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 western blot western blot Immuno blot Immuno blot Beinke S., Deka J., Lang V., Belich M.P., Walker P.A., Howell S., Smerdon S.J., Gamblin S.J., Ley S.C. IBD - Inflammatory bowel disease TPL2 constructs are described in PMID:9950430; p105 constructs are described in PMID:11976329 and PMID:11297557. Information about GST-tagged inactive MEK1 was retrieved from the suppliers website (Upstate Biotechnology). sley@nimr.mrc.ac.uk,ssmerdo@nimr.mrc.ac.uk Mol. Cell. Biol. (0270-7306) 2003 Only protein-protein interactions imex curation beinke-2003-8 NF-kappaB1 p105 negatively regulates TPL-2 MEK kinase activity. in vitro In vitro cosedimentation cosedimentation predetermined predetermined participant predetermined Beinke S., Deka J., Lang V., Belich M.P., Walker P.A., Howell S., Smerdon S.J., Gamblin S.J., Ley S.C. IBD - Inflammatory bowel disease TPL2 constructs are described in PMID:9950430; p105 constructs are described in PMID:11976329 and PMID:11297557. Information about GST-tagged inactive MEK1 was retrieved from the suppliers website (Upstate Biotechnology). sley@nimr.mrc.ac.uk,ssmerdo@nimr.mrc.ac.uk Mol. Cell. Biol. (0270-7306) 2003 Only protein-protein interactions imex curation beinke-2003-9 NF-kappaB1 p105 negatively regulates TPL-2 MEK kinase activity. in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Beinke S., Deka J., Lang V., Belich M.P., Walker P.A., Howell S., Smerdon S.J., Gamblin S.J., Ley S.C. IBD - Inflammatory bowel disease TPL2 constructs are described in PMID:9950430; p105 constructs are described in PMID:11976329 and PMID:11297557. Information about GST-tagged inactive MEK1 was retrieved from the suppliers website (Upstate Biotechnology). sley@nimr.mrc.ac.uk,ssmerdo@nimr.mrc.ac.uk Mol. Cell. Biol. (0270-7306) 2003 Only protein-protein interactions imex curation beinke-2003-2 NF-kappaB1 p105 negatively regulates TPL-2 MEK kinase activity. in vitro In vitro pull down pull down autoradiography autoradiography Beinke S., Deka J., Lang V., Belich M.P., Walker P.A., Howell S., Smerdon S.J., Gamblin S.J., Ley S.C. IBD - Inflammatory bowel disease TPL2 constructs are described in PMID:9950430; p105 constructs are described in PMID:11976329 and PMID:11297557. Information about GST-tagged inactive MEK1 was retrieved from the suppliers website (Upstate Biotechnology). sley@nimr.mrc.ac.uk,ssmerdo@nimr.mrc.ac.uk Mol. Cell. Biol. (0270-7306) 2003 Only protein-protein interactions imex curation beinke-2003-3 NF-kappaB1 p105 negatively regulates TPL-2 MEK kinase activity. in vitro In vitro spr surface plasmon resonance Optical biosensor BIAcore(r) BIAcore(r) Optical biosensor spr predetermined predetermined participant predetermined Beinke S., Deka J., Lang V., Belich M.P., Walker P.A., Howell S., Smerdon S.J., Gamblin S.J., Ley S.C. IBD - Inflammatory bowel disease TPL2 constructs are described in PMID:9950430; p105 constructs are described in PMID:11976329 and PMID:11297557. Information about GST-tagged inactive MEK1 was retrieved from the suppliers website (Upstate Biotechnology). sley@nimr.mrc.ac.uk,ssmerdo@nimr.mrc.ac.uk Mol. Cell. Biol. (0270-7306) 2003 Only protein-protein interactions imex curation beinke-2003-4 NF-kappaB1 p105 negatively regulates TPL-2 MEK kinase activity. in vitro In vitro itc isothermal titration calorimetry ITC ITC itc predetermined predetermined participant predetermined Beinke S., Deka J., Lang V., Belich M.P., Walker P.A., Howell S., Smerdon S.J., Gamblin S.J., Ley S.C. IBD - Inflammatory bowel disease TPL2 constructs are described in PMID:9950430; p105 constructs are described in PMID:11976329 and PMID:11297557. Information about GST-tagged inactive MEK1 was retrieved from the suppliers website (Upstate Biotechnology). sley@nimr.mrc.ac.uk,ssmerdo@nimr.mrc.ac.uk Mol. Cell. Biol. (0270-7306) 2003 Only protein-protein interactions imex curation beinke-2003-5 NF-kappaB1 p105 negatively regulates TPL-2 MEK kinase activity. in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Beinke S., Deka J., Lang V., Belich M.P., Walker P.A., Howell S., Smerdon S.J., Gamblin S.J., Ley S.C. IBD - Inflammatory bowel disease TPL2 constructs are described in PMID:9950430; p105 constructs are described in PMID:11976329 and PMID:11297557. Information about GST-tagged inactive MEK1 was retrieved from the suppliers website (Upstate Biotechnology). sley@nimr.mrc.ac.uk,ssmerdo@nimr.mrc.ac.uk Mol. Cell. Biol. (0270-7306) 2003 Only protein-protein interactions imex curation beinke-2003-6 NF-kappaB1 p105 negatively regulates TPL-2 MEK kinase activity. in vitro In vitro enzymatic footprint enzymatic footprinting enzymatic footprint edman degradation edman degradation Beinke S., Deka J., Lang V., Belich M.P., Walker P.A., Howell S., Smerdon S.J., Gamblin S.J., Ley S.C. IBD - Inflammatory bowel disease TPL2 constructs are described in PMID:9950430; p105 constructs are described in PMID:11976329 and PMID:11297557. Information about GST-tagged inactive MEK1 was retrieved from the suppliers website (Upstate Biotechnology). sley@nimr.mrc.ac.uk,ssmerdo@nimr.mrc.ac.uk Mol. Cell. Biol. (0270-7306) 2003 Only protein-protein interactions imex curation nfkb1_human Nuclear factor NF-kappa-B p105 subunit NFKB1 DNA-binding factor KBF1 EBP-1 Nuclear factor of kappa light polypeptide gene enhancer in B-cells 1 protein protein human Homo sapiens Human MAEDDPYLGRPEQMFHLDPSLTHTIFNPEVFQPQMALPTDGPYLQILEQPKQRGFRFRYVCEGPSHGGLPGASSEKNKKSYPQVKICNYVGPAKVIVQLVTNGKNIHLHAHSLVGKHCEDGICTVTAGPKDMVVGFANLGILHVTKKKVFETLEARMTEACIRGYNPGLLVHPDLAYLQAEGGGDRQLGDREKELIRQAALQQTKEMDLSVVRLMFTAFLPDSTGSFTRRLEPVVSDAIYDSKAPNASNLKIVRMDRTAGCVTGGEEIYLLCDKVQKDDIQIRFYEEEENGGVWEGFGDFSPTDVHRQFAIVFKTPKYKDINITKPASVFVQLRRKSDLETSEPKPFLYYPEIKDKEEVQRKRQKLMPNFSDSFGGGSGAGAGGGGMFGSGGGGGGTGSTGPGYSFPHYGFPTYGGITFHPGTTKSNAGMKHGTMDTESKKDPEGCDKSDDKNTVNLFGKVIETTEQDQEPSEATVGNGEVTLTYATGTKEESAGVQDNLFLEKAMQLAKRHANALFDYAVTGDVKMLLAVQRHLTAVQDENGDSVLHLAIIHLHSQLVRDLLEVTSGLISDDIINMRNDLYQTPLHLAVITKQEDVVEDLLRAGADLSLLDRLGNSVLHLAAKEGHDKVLSILLKHKKAALLLDHPNGDGLNAIHLAMMSNSLPCLLLLVAAGADVNAQEQKSGRTALHLAVEHDNISLAGCLLLEGDAHVDSTTYDGTTPLHIAAGRGSTRLAALLKAAGADPLVENFEPLYDLDDSWENAGEDEGVVPGTTPLDMATSWQVFDILNGKPYEPEFTSDDLLAQGDMKQLAEDVKLQLYKLLEIPDPDKNWATLAQKLGLGILNNAFRLSPAPSKTLMDNYEVSGGTVRELVEALRQMGYTEAIEVIQAASSPVKTTSQAHSLPLSPASTRQQIDELRDSDSVCDSGVETSFRKLSFTESLTSGASLLTLNKMPHDYGQEGPLEGKI m3k8_human Mitogen-activated protein kinase kinase kinase 8 MAP3K8 COT ESTF Serine/threonine-protein kinase cot Cancer Osaka thyroid oncogene Tumor progression locus 2 Proto-oncogene c-Cot protein protein human Homo sapiens Human MEYMSTGSDNKEEIDLLIKHLNVSDVIDIMENLYASEEPAVYEPSLMTMCQDSNQNDERSKSLLLSGQEVPWLSSVRYGTVEDLLAFANHISNTAKHFYGQRPQESGILLNMVITPQNGRYQIDSDVLLIPWKLTYRNIGSDFIPRGAFGKVYLAQDIKTKKRMACKLIPVDQFKPSDVEIQACFRHENIAELYGAVLWGETVHLFMEAGEGGSVLEKLESCGPMREFEIIWVTKHVLKGLDFLHSKKVIHHDIKPSNIVFMSTKAVLVDFGLSVQMTEDVYFPKDLRGTEIYMSPEVILCRGHSTKADIYSLGATLIHMQTGTPPWVKRYPRSAYPSYLYIIHKQAPPLEDIADDCSPGMRELIEASLERNPNHRPRAADLLKHEALNPPREDQPRCQSLDSALLERKRLLSRKELELPENIADSSCTGSTEESEMLKRQRSLYIDLGALAGYFNLVRGPPTLEYG 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 nfkb1-map3k8 1573668 n/a 1573676 unspecified role unspecified role bait bait n-terminus 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 false n/a 1573677 unspecified role unspecified role prey prey prot_kinase_dom sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false n-terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false P41279:p.Asp270Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Fig. 1 nfkb1-map3k8-2 1573668 n/a 1573677 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n-terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false n/a 1573676 unspecified role unspecified role bait bait P19838:p.Leu841Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false n-terminus 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 false death_domain required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Fig. 2b nfkb1-map3k8-3 1573668 n/a 1573676 unspecified role unspecified role bait bait region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false P19838:p.Leu841Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false n-terminus 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 false n/a 1573677 unspecified role unspecified role prey prey physical association physical association false false false Deletion of the p105 death domain (residues 802-892) or p105 residues 497-538 separately decreased binding to TPL-2, while combined deletion of both these regions in p105 disrupted the interaction. Fig. 5c When combined with deletion of p105 residues 497-538, the leu841ala mutation disrupts binding to TPL-2. nfkb1-nfkb1-5 1573669 n/a 1573676 unspecified role unspecified role neutral component neutral component region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false ecobd Escherichia coli (strain B / BL21-DE3) Stoichiometry: 1.0 n/a 1573676 unspecified role unspecified role neutral component neutral component region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false ecobd Escherichia coli (strain B / BL21-DE3) Stoichiometry: 1.0 1573704 1573702 direct interaction direct interaction false false false Fig. 4b map2k1-map3k8-3 1573670 n/a 1573678 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 ecolx Escherichia coli n/a 1573677 unspecified role unspecified role prey prey n-terminus myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false 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. physical association physical association false false false Fig. 9b, 9c P19838 nfkb1-map3k8-1 1573671 n/a 1573677 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false in vitro In vitro n/a 1573676 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 death_domain sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false ecobd Escherichia coli (strain B / BL21-DE3) direct interaction direct interaction false false false Fig. 2c map3k8-nfkb1-1 1573672 n/a 1573677 unspecified role unspecified role bait bait n-terminus biotin tag biotin tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1573676 unspecified role unspecified role prey prey region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false ecobd Escherichia coli (strain B / BL21-DE3) direct interaction direct interaction false false false 1573672 Fig. 3c, 3d Temp = 25C The C-terminal fragment of TPL-2 (residues 398-467) bound to the p105 construct (residues 497-968) with Kd = 138 x 10E-9 molar, to p105 (residues 497-805) with Kd = 123 x 10E-9 molar and to p105 (residues 497-782) with Kd = 88 X 10E-9 molar. No binding to p105 (residues 540-801) was detected. map3k8-nfkb1-2 1573673 n/a 1573677 unspecified role unspecified role neutral component neutral component n-terminus biotin tag biotin tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) Stoichiometry: 1.0 n/a 1573676 unspecified role unspecified role neutral component neutral component region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false ecobd Escherichia coli (strain B / BL21-DE3) Stoichiometry: 2.0 1573727 1573725 direct interaction direct interaction false false false 1573673 P19838 decreases degradation at protein level of P41279 P19838 decreases activity of P41279 P19838 decreases binding of P41279 to Q02750 Fig. 4a Temp = 20C The Kd of 58 x 10E-9 molar was determined for binding of the C-terminal region of TPL-2 (residues 398-467) to p105 construct (residues 497-805). TPL-2 (residues 398-467) bound to p105 (residues 497-757) with Kd = 64 x 10E-9 molar. No binding was detected to p105 (residues 540-801). map3k8-nfkb1-3 1573674 n/a 1573677 unspecified role unspecified role bait bait c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n-terminus biotin tag biotin tag 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 1573676 unspecified role unspecified role prey prey n-terminus 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 false region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false 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. physical association physical association false false false Fig. 5b map3k8-nfkb1-4 1573675 n/a 1573677 unspecified role unspecified role neutral component neutral component n-terminus biotin tag biotin tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false c-terminal binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) n/a 1573676 unspecified role unspecified role neutral component neutral component ecobd Escherichia coli (strain B / BL21-DE3) direct interaction direct interaction false false false Fig. 3b