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/ tu-2010-2 DAPK1 Interaction with NMDA Receptor NR2B Subunits Mediates Brain Damage in Stroke. mouse-brain-cortex Mus musculus brain cortex brain_cortex Brain cortex [Cerebral cortex; Cerebrum cortex; Cortex; Pallium] anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Tu W., Xu X., Peng L., Zhong X., Zhang W., Soundarapandian MM., Balel C., Wang M., Jia N., Zhang W., Lew F., Chan SL., Chen Y., Lu Y. Apoptosis - Interactions involving proteins with a function related to apoptosis ylu1@lsuhsc.edu Cell (0092-8674) 2010 Only protein-protein interactions 9 imex curation Synapse - Interactions of proteins with an established role in the presynapse. tu-2010-4 DAPK1 Interaction with NMDA Receptor NR2B Subunits Mediates Brain Damage in Stroke. in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Tu W., Xu X., Peng L., Zhong X., Zhang W., Soundarapandian MM., Balel C., Wang M., Jia N., Zhang W., Lew F., Chan SL., Chen Y., Lu Y. Apoptosis - Interactions involving proteins with a function related to apoptosis Source of expression constructs supplied by the author. Details of expression systems are not available. Data from Fig. S1 not added as source of NR1 not available. ylu1@lsuhsc.edu Cell (0092-8674) 2010 Only protein-protein interactions 9 imex curation Synapse - Interactions of proteins with an established role in the presynapse. tu-2010-1 DAPK1 Interaction with NMDA Receptor NR2B Subunits Mediates Brain Damage in Stroke. mouse-brain-cortex Mus musculus brain cortex brain_cortex Brain cortex [Cerebral cortex; Cerebrum cortex; Cortex; Pallium] anti bait coip anti bait coimmunoprecipitation anti bait coip fingerprinting peptide massfingerprinting fingerprinting Tu W., Xu X., Peng L., Zhong X., Zhang W., Soundarapandian MM., Balel C., Wang M., Jia N., Zhang W., Lew F., Chan SL., Chen Y., Lu Y. Apoptosis - Interactions involving proteins with a function related to apoptosis ylu1@lsuhsc.edu Cell (0092-8674) 2010 Only protein-protein interactions 9 imex curation Synapse - Interactions of proteins with an established role in the presynapse. tu-2010-3 DAPK1 Interaction with NMDA Receptor NR2B Subunits Mediates Brain Damage in Stroke. mouse-neuron Mus musculus neuronal cell neuron Neuron [Neuronal cell; Neural cell fluorescence imaging fluorescence microscopy fluorescence imaging immunostaining immunostaining Tu W., Xu X., Peng L., Zhong X., Zhang W., Soundarapandian MM., Balel C., Wang M., Jia N., Zhang W., Lew F., Chan SL., Chen Y., Lu Y. Apoptosis - Interactions involving proteins with a function related to apoptosis ylu1@lsuhsc.edu Cell (0092-8674) 2010 Only protein-protein interactions 9 imex curation Synapse - Interactions of proteins with an established role in the presynapse. nmdz1_mouse Glutamate receptor ionotropic, NMDA 1 Glutamate [NMDA] receptor subunit zeta-1 Grin1 Glurz1 N-methyl-D-aspartate receptor subunit NR1 protein protein mouse Mus musculus Mouse MSTMHLLTFALLFSCSFARAACDPKIVNIGAVLSTRKHEQMFREAVNQANKRHGSWKIQLNATSVTHKPNAIQMALSVCEDLISSQVYAILVSHPPTPNDHFTPTPVSYTAGFYRIPVLGLTTRMSIYSDKSIHLSFLRTVPPYSHQSSVWFEMMRVYNWNHIILLVSDDHEGRAAQKRLETLLEERESKAEKVLQFDPGTKNVTALLMEARDLEARVIILSASEDDAATVYRAAAMLNMTGSGYVWLVGEREISGNALRYAPDGIIGLQLINGKNESAHISDAVGVVAQAVHELLEKENITDPPRGCVGNTNIWKTGPLFKRVLMSSKYADGVTGRVEFNEDGDRKFANYSIMNLQNRKLVQVGIYNGTHVIPNDRKIIWPGGETEKPRGYQMSTRLKIVTIHQEPFVYVKPTMSDGTCKEEFTVNGDPVKKVICTGPNDTSPGSPRHTVPQCCYGFCVDLLIKLARTMNFTYEVHLVADGKFGTQERVNNSNKKEWNGMMGELLSGQADMIVAPLTINNERAQYIEFSKPFKYQGLTILVKKEIPRSTLDSFMQPFQSTLWLLVGLSVHVVAVMLYLLDRFSPFGRFKVNSEEEEEDALTLSSAMWFSWGVLLNSGIGEGAPRSFSARILGMVWAGFAMIIVASYTANLAAFLVLDRPEERITGINDPRLRNPSDKFIYATVKQSSVDIYFRRQVELSTMYRHMEKHNYESAAEAIQAVRDNKLHAFIWDSAVLEFEASQKCDLVTTGELFFRSGFGIGMRKDSPWKQNVSLSILKSHENGFMEDLDKTWVRYQECDSRSNAPATLTFENMAGVFMLVAGGIVAGIFLIFIEIAYKRHKDARRKQMQLAFAAVNVWRKNLQDRKSGRAEPDPKKKATFRAITSTLASSFKRRRSSKDTSTGGGRGALQNQKDTVLPRRAIEREEGQLQLCSRHRES dlg4_mouse Disks large homolog 4 Dlg4 Dlgh4 Psd95 Postsynaptic density protein 95 Synapse-associated protein 90 protein protein mouse Mus musculus Mouse MDCLCIVTTKKYRYQDEDTPPLEHSPAHLPNQANSPPVIVNTDTLEAPGYELQVNGTEGEMEYEEITLERGNSGLGFSIAGGTDNPHIGDDPSIFITKIIPGGAAAQDGRLRVNDSILFVNEVDVREVTHSAAVEALKEAGSIVRLYVMRRKPPAEKIIEIKLIKGPKGLGFSIAGGVGNQHIPGDNSIYVTKIIEGGAAHKDGRLQIGDKILAVNSVGLEDVMHEDAVAALKNTYDVVYLKVAKPSNAYLSDSYAPPDITTSYSQHLDNEISHSSYLGTDYPTAMTPTSPRRYSPVAKDLLGEEDIPREPRRIVIHRGSTGLGFNIVGGEDGEGIFISFILAGGPADLSGELRKGDQILSVNGVDLRNASHEQAAIALKNAGQTVTIIAQYKPEEYSRFEAKIHDLREQLMNSSLGSGTASLRSNPKRGFYIRALFDYDKTKDCGFLSQALSFHFGDVLHVIDASDEEWWQARRVHSDSETDDIGFIPSKRRVERREWSRLKAKDWGSSSGSQGREDSVLSYETVTQMEVHYARPIIILGPTKDRANDDLLSEFPDKFGSCVPHTTRPKREYEIDGRDYHFVSSREKMEKDIQAHKFIEAGQYNSHLYGTSVQSVREVAEQGKHCILDVSANAVRRLQAAHLHPIAIFIRPRSLENVLEINKRITEEQARKAFDRATKLEQEFTECFSAIVEGDSFEEIYHKVKRVIEDLSGPYIWVPARERL nmde2_mouse Glutamate receptor ionotropic, NMDA 2B Glutamate [NMDA] receptor subunit epsilon-2 Grin2b N-methyl D-aspartate receptor subtype 2B protein protein mouse Mus musculus Mouse MKPSAECCSPKFWLVLAVLAVSGSKARSQKSAPSIGIAVILVGTSDEVAIKDAHEKDDFHHLSVVPRVELVAMNETDPKSIITRICDLMSDRKIQGVVLADDTDQEAIAQILDFISAQTLTPILGIHGGSSMIMADKDESSMFFQFGPSIEQQASVMLNIMEEYDWYIFSIVTTYFPGYQDFVNKIRSTIENSFVGWELEEVLLLDMSLDDGDSKIQNQLKKLQSPIILLYCTKEEATYIFEVANSVGLTGYGYTWIVPSLVAGDTDTVPSEFPTGLISVSYDEWDYGLPARVRDGIAIITTAASDMLSEHSFIPEPKSSCYNTHEKRIYQSNMLNRYLINVTFEGRNLSFSEDGYQMHPKLVIILLNKERKWERVGKWKDKSLQMKYYVWPRMCPETEEQEDDHLSIVTLEEAPFVIVESVDPLSGTCMRNTVPCQKRIISENKTDEEPGYIKKCCKGFCIDILKKISKSVKFTYDLYLVTNGKHGKKINGTWNGMIGEVVMKRAYMAVGSLTINEERSEVVDFSVPFIETGISVMVSRSNGTVSPSAFLEPFSADVWVMMFVMLLIVSAVAVFVFEYFSPVGYNRCLADGREPGGPSFTIGKAIWLLWGLVFNNSVPVQNPKGTTSKIMVSVWAFFAVIFLASYTANLAAFMIQEEYVDQVSGLSDKKFQRPNDFSPPFRFGTVPNGSTERNIRNNYAEMHAYMGKFNQRGVDDALLSLKTGKLDAFIYDAAVLNYMAGRDEGCKLVTIGSGKVFASTGYGIAIQKDSGWKRQVDLAILQLFGDGEMEELEALWLTGICHNEKNEVMSSQLDIDNMAGVFYMLGAAMALSLITFICEHLFYWQFRHCFMGVCSGKPGMVFSISRGIYSCIHGVAIEERQSVMNSPTATMNNTHSNILRLLRTAKNMANLSGVNGSPQSALDFIRRESSVYDISEHRRSFTHSDCKSYNNPPCEENLFSDYISEVERTFGNLQLKDSNVYQDHYHHHHRPHSIGSTSSIDGLYDCDNPPFTTQPRSISKKPLDIGLPSSKHSQLSDLYGKFSFKSDRYSGHDDLIRSDVSDISTHTVTYGNIEGNAAKRRKQQYKDSLKKRPASAKSRREFDEIELAYRRRPPRSPDHKRYFRDKEGLRDFYLDQFRTKENSPHWEHVDLTDIYKERSDDFKRDSVSGGGPCTNRSHLKHGTGDKHGVVGGVPAPWEKNLTNVDWEDRSGGNFCRSCPSKLHNYSSTVAGQNSGRQACIRCEACKKAGNLYDISEDNSLQELDQPAAPVAVSSNASTTKYPQSPTNSKAQKKNRNKLRRQHSYDTFVDLQKEEAALAPRSVSLKDKGRFMDGSPYAHMFEMPAGESSFANKSSVTTAGHHHNNPGSGYMLSKSLYPDRVTQNPFIPTFGDDQCLLHGSKSYFFRQPTVAGASKTRPDFRALVTNKPVVSALHGAVPGRFQKDICIGNQSNPCVPNNKNPRAFNGSSNGHVYEKLSSIESDV dapk1_mouse Death-associated protein kinase 1 Dapk1 protein protein mouse Mus musculus Mouse MTVFRQENVDDYYDTGEELGSGQFAVVKKCREKSTGLQYAAKFIKKRRTKSSRRGVSREDIEREVSILKEIRHPNVITLHEVYENKTDVILILELVAGGELFDFLAEKESLTEEEATEFLKQILSGVYYLHSLQIAHFDLKPENIMLLDRNVPKPRIKIIDFGLAHKIDFGNEFKNIFGTPEFVAPEIVNYEPLGLEADMWSIGVITYILLSGASPFLGDTKQETLANVSAVNYDFEEEFFRNTSTLAKDFIRRLLVKDPKKRMTIQDSLQHPWIKPKDTQQALSRKASAVNMEKFKKFAARKKWKQSVRLISLCQRLSRSFLSRSNMSVARSDDTLDEEDSFVMKAIIHAINDDNVPGLQHLLGSLSSYDVNQPNKHGTPPLLIAAGCGNIQMLQLLIKRGSRIDVQDKGGSNAIYWASRHGHVDTLKFLNENKCPLDVKDKSGETALHVAARYGHADVVQLLCSFGSNPDFQDKEEETPLHCAAWHGYYSVAKALCEVGCNVNIKNREGETPLLTASARGYHDIVECLAEHGADLNASDKDGHIALHLAVRRCQMEVIKTLLGHGSFVDFQDRHGNTPLHVACKDGSAPIVVALCEASCNLDISNKYGRTPLHLAANNGILDVVRYLCLMGANVEALTSDGKTAEDLAKAEQHEHVAGLLARLRKDTHRGLFIQQLRPTQNLQPRIKLKLFGHSGSGKSTLVESLKCGLLRSFFRRRRPRLSSTNSTRFPPSPLAAKPTVSVSINNLYPGCENVSVRSRSMMFEPGLTKGMLEVFVAPSHHLHCSTDDQSTKAIDIQNAYLNGVGDFSVWEFSGNPVYFCCYDYFAANDPTSIHIIVFSLEEPYEIQLNQVIFWLSFLKSLVPVEEPIAFGGKLKNPLRVVLVATHADIMNIPRPAGGEFGYDKDTSLLKEIRNRFGNDLHVSNKLFVLDAGASGSKDIKVLRNHLQEIRSQIVSGCSPMTHLCEKIISTLPSWRKLNGPNQLMSLQQFVYDVQDQLNPLASEDDLRRIAQQLHSTGEINIMQSETVQDVLLLDPRWLCTNVLGKLLSVETPRALHHYRGRYTMEDIQRLVPDSDVEELLQILDAMDICARDLSSGTMVDIPALIKTDSLQRSWADEEDEVMVYGGVRIVPVEHLTPFPCGIFHKVQVNLCRWIHQQSAEGDADIRLWVSGCRIANRGAELLVLLVNHGQGIEVQVRGLETEKIKCCLLLDSVCSTIETVMATTLPGLLTVKHYLSPQQLREHHEPVMVYQPRDFFRAQTLKESSLTNTMGGYKESFSSITCFGCHDVYSQASLGMDIHASDLSLLTRRKLSRLLDPPDPMGKDWCLLAMNLGLPDMVAKHNVNNRASRDFLPSPVHALLQEWTSYPESTVGILISKLRELGRRDAADFLLKASSVFKINLDGNGQEAYASSCNSGTSYNSISSVVSRRDSHAWTPLYDL kcc2a_mouse Calcium/calmodulin-dependent protein kinase type II subunit alpha Camk2a protein protein mouse Mus musculus Mouse MATITCTRFTEEYQLFEELGKGAFSVVRRCVKVLAGQEYAAKIINTKKLSARDHQKLEREARICRLLKHPNIVRLHDSISEEGHHYLIFDLVTGGELFEDIVAREYYSEADASHCIQQILEAVLHCHQMGVVHRDLKPENLLLASKLKGAAVKLADFGLAIEVEGEQQAWFGFAGTPGYLSPEVLRKDPYGKPVDLWACGVILYILLVGYPPFWDEDQHRLYQQIKAGAYDFPSPEWDTVTPEAKDLINKMLTINPSKRITAAEALKHPWISHRSTVASCMHRQETVDCLKKFNARRKLKGAILTTMLATRNFSGGKSGGNKKNDGVKESSESTNTTIEDEDTKVRKQEIIKVTEQLIEAISNGDFESYTKMCDPGMTAFEPEALGNLVEGLDFHRFYFENLWSRNSKPVHTTILNPHIHLMGDESACIAYIRITQYLDAGGIPRTAQSEETRVWHRRDGKWQIVHFHRSGAPSVLPH grin2b-dapk1-2 Interactors of Grin2b (Nr2b) - ischemic mice 883390 n/a 883394 unspecified role unspecified role prey prey n/a 883395 unspecified role unspecified role prey prey n/a 883396 unspecified role unspecified role bait bait n/a 883397 unspecified role unspecified role prey prey physical association physical association false false false Interaction observed in mice operated on to induce ischemia. Fig. 1C grin2b-grin1-1 Interactors of Grin2b (Nr2b) - non-ischemic mice 883390 n/a 883395 unspecified role unspecified role prey prey n/a 883394 unspecified role unspecified role prey prey n/a 883396 unspecified role unspecified role bait bait physical association physical association false false false Interaction observed in non-ischemic mice. Fig. 1C grin2b-dapk1-3 Interactors of Dapk1 - ischemic mice 883390 n/a 883397 unspecified role unspecified role bait bait n/a 883395 unspecified role unspecified role prey prey n/a 883396 unspecified role unspecified role prey prey n/a 883394 unspecified role unspecified role prey prey physical association physical association false false false Interaction observed in mice operated on to induce ischemia. Fig. 1C grin2b-dapk1-7 Interaction between Grin2b (Nr2b) and DAPK1 demonstrated by coimmunoprecipitation 883390 n/a 883396 unspecified role unspecified role bait bait n/a 883397 unspecified role unspecified role prey prey n/a 883395 unspecified role unspecified role prey prey n/a 883398 unspecified role unspecified role prey prey association association false false false Membrane extracts were precipitated with nonspecific IgG (IgG) or anti-NR2B in the presence of 50 mM NR2BCT or sNR2BCT and probed with anti-DAPK1, or anti-PSD-95, or anti-CaMK-IIa. Fig. 2D When immunoprecipitation were performed in the presence of Nr2b C-terminal peptide, the interaction with Dak1 but not Dlg4 or Camk2a was disrupted. grin2b-dapk1-8 Interaction between Grin2b (Nr2b) and DAPK1 demonstrated by coimmunoprecipitation 883390 n/a 883396 unspecified role unspecified role bait bait n/a 883397 unspecified role unspecified role prey prey physical association physical association false false false Anti-NR2B precipitates were blotted with anti-DAPK1. Supp. Fig. 2E NMDA/glycine - activates endogenous DAPK1. grin2b-dapk1-5 Interaction between Grin2b (Nr2b) and DAPK1 demonstrated by pulldown 883391 n/a 883396 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 false 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 n/a 883397 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 false catalytic region sufficient to bind sufficient binding region sufficient to bind range range range range false 883427 883423 direct interaction direct interaction false false false Affinity binding assay shows that flag-tagged catalytic domain of DAPK1 binds with GST-NR2B839-1482 (∼100 kDa) and GST-NR2B1292-1304 (∼30 kDa). The blots show anti-Flag (top) and anti-GST (bottom). Figs. 2A, 2B grin2b-dapk1-6 Interaction between Grin2b (Nr2b) and DAPK1 demonstrated by pulldown 883391 n/a 883396 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 false 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 n/a 883396 competitor competitor competitor prey prey region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false in vitro In vitro n/a 883397 unspecified role unspecified role prey prey mouse-brain Mus musculus brain brain Brain [cerebrum] association association false false false Affinity binding assay shows that flag-tagged catalytic domain of DAPK1 binds with GST-NR2B839-1482 (∼100 kDa) and GST-NR2B1292-1304 (∼30 kDa). The blots show anti-Flag (top) and anti-GST (bottom). Figs. 2A, 2B grin2b-dapk1-1 Interaction between Grin2b (Nr2b) and Dapk1 demontrated by coimmunoprecipitation 883392 n/a 883397 unspecified role unspecified role prey prey n/a 883396 unspecified role unspecified role bait bait physical association physical association false false false Interaction observed in mice operated on to induce ischemia. Figs. 1 and 1B grin2b-dapk1-4 Colocalisation of Grin2b (Nr2b) and Dapk1 883393 n/a 883396 unspecified role unspecified role neutral component neutral component n/a 883397 unspecified role unspecified role neutral component neutral component colocalization colocalization false false false Figs. 1E and 1F