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/ Cordon-Bleu Is an Actin Nucleation Factor and Controls Neuronal Morphology. Cordon-Bleu Is an Actin Nucleation Factor and Controls Neuronal Morphology. Cell (0092-8674) 2007 imex curation Ahuja R., Pinyol R., Reichenbach N., Custer L., Klingensmith J., Kessels MM., Qualmann B. Synapse - Interactions of proteins with an established role in the presynapse. Source of Syndactin i (Pacsin1) taken form PMID: 9950691 qualmann@ifn-magdeburg.de 22-Jan-2008: Contacted by JYOTI. Mouse brain cDNA DAVET Only protein-protein interactions 6 in vitro In vitro pull down pull down tag fluorescence tag visualisation by fluorescence tag fluorescence Ahuja R., Pinyol R., Reichenbach N., Custer L., Klingensmith J., Kessels MM., Qualmann B. Synapse - Interactions of proteins with an established role in the presynapse. Interactions with actin identified only by antibody staining could not be added due to non-specificity of Abs for different actin gene products Beads coated with recombinant proteins were incubated with rat brain extracts supplemented with an energy regenerating mix and fluorescently labeled rabbit skeletal G-actin. Accumulation of labelled actim polmerizing on the bead surface was taken as a measure of interaction with the bait protein Source of Syndactin i (Pacsin1) taken form PMID: 9950691 qualmann@ifn-magdeburg.de qualmann@ifn-magdeburg.de Accepted 2007-DEC-20 by DAVET. 22-Jan-2008: Contacted by JYOTI. Mouse brain cDNA Cell (0092-8674) 2007 DAVET Only protein-protein interactions 6 imex curation Cordon-Bleu Is an Actin Nucleation Factor and Controls Neuronal Morphology. Cordon-Bleu Is an Actin Nucleation Factor and Controls Neuronal Morphology. Cell (0092-8674) 2007 imex curation Ahuja R., Pinyol R., Reichenbach N., Custer L., Klingensmith J., Kessels MM., Qualmann B. Synapse - Interactions of proteins with an established role in the presynapse. Source of Syndactin i (Pacsin1) taken form PMID: 9950691 qualmann@ifn-magdeburg.de 22-Jan-2008: Contacted by JYOTI. Mouse brain cDNA DAVET Only protein-protein interactions 6 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 Ahuja R., Pinyol R., Reichenbach N., Custer L., Klingensmith J., Kessels MM., Qualmann B. Synapse - Interactions of proteins with an established role in the presynapse. Source of Syndactin i (Pacsin1) taken form PMID: 9950691 qualmann@ifn-magdeburg.de qualmann@ifn-magdeburg.de Accepted 2007-DEC-20 by DAVET. 22-Jan-2008: Contacted by JYOTI. Mouse brain cDNA Cell (0092-8674) 2007 DAVET Only protein-protein interactions 6 imex curation Cordon-Bleu Is an Actin Nucleation Factor and Controls Neuronal Morphology. Cordon-Bleu Is an Actin Nucleation Factor and Controls Neuronal Morphology. Cell (0092-8674) 2007 imex curation Ahuja R., Pinyol R., Reichenbach N., Custer L., Klingensmith J., Kessels MM., Qualmann B. Synapse - Interactions of proteins with an established role in the presynapse. Source of Syndactin i (Pacsin1) taken form PMID: 9950691 qualmann@ifn-magdeburg.de 22-Jan-2008: Contacted by JYOTI. Mouse brain cDNA DAVET Only protein-protein interactions 6 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 Ahuja R., Pinyol R., Reichenbach N., Custer L., Klingensmith J., Kessels MM., Qualmann B. Synapse - Interactions of proteins with an established role in the presynapse. Source of Syndactin i (Pacsin1) taken form PMID: 9950691 qualmann@ifn-magdeburg.de qualmann@ifn-magdeburg.de Accepted 2007-DEC-20 by DAVET. Mouse brain cDNA Cell (0092-8674) 2007 DAVET Only protein-protein interactions 6 imex curation cobl_mouse Protein cordon-bleu Cobl Kiaa0633 protein protein mouse Mus musculus Mouse MDAPRALAAKPPTGRKMKARAPPPPGKPAAQNVHSEQKLPHDATLGSQQSLVYMKEALQNSTLDITVVLPSGLEKQSVVSGSHAMMDLLVELCLQNHLNPSHHVLEIWSSETQQPLSFKPNTLIGSLNVHTVLLKEKVPEERVKPGLTKAPEKSVRLVVNYLRTQKAVVRVSPEVPLQNILPVICAKCEVNPEHVILLRDNVAGEELELSKSLNELGIKELYAWDNRREMFRKSSLGNDETDKEKKKFLGFFKANKRSNSKAEHLGLSGADSDEDPAKSASGGDLNGCVTTPNSPSLHSRSLTLGPSLSLGNISGVSMKSDMKKRRAPPPPSPKLLGQDKVSEKASLSSQADLQKKKRRAPAPPPPQQPPPSPVVPNRKEDKEENRKSTVGVGRQVPQKPPRGTARGPPQLVLPPPPPYPPPDTDVTEPVTFPGEGAGSETSELRPKLSLPLGPGSHCSMGGVSQVPAESEETASEDTTEDSGVMSSPSDAISLDSQQDSMRSKDKWSTDQEDGSDQDLAGTPELGPQKSPSWGKSGSGSSILRTEKATMPTNDDEDLFITGHLHQTLAELDEDLEGMEENYETDTSSLTNSVNGVSNHSLQEAIIPDSGVDDIPVTFIGEVSDEPFDSGLFSSRCNNATTFNTGSIASQRSHLSPSQTEHSQPFVRTSRKEPDPSPPSQDNRKRNQPTLANTSENENPVETDPTVTSLVSKLLIDDPKAKDKGKVHGSSHSEKTQAGHGINSLRVNPRDGKDESSNSAPPPWSHHGQALGGSYGLKYGLTTYKIVPPKSEMRCYDRDVSLSTGAIKIDELGNLVSPHMNGSRTISPPSAVVETDTPPIGKVKEFWRRNSMEKYLNGPAECTIKRAPSTTITATPEKPQQDNGMKAAFTVTTPQQQPASQEYGAHLEEERSRPQSAVSCSVKVPASNPTDITFLKPQRRTSSQYVASAIAKKMGPPKVHADVVRPHKATTEQCHEEAKLARSPPTRKDDAAPNLHSEARQHEHGTNQSSVCLPSNPGVQLPAGGHPKVEVNSTYGKSSTQDYPAAVHRNSYFLPGRSSHRDRVSVGQSCGFNEKQTTSNQKANSTSNFSQALDKAHPPPLLLAEARDSGRILMNGSARTPGNCEPPHSPKESTLTSYIILQTEEKPSSLSTDGQDADDTLPSSIFGPKKKFKPVIQRPLPKDVSLHSALMEAIHSSGGREKLRKTAEQTSEGRPKKPSYVEAESERSALLAAIRGHSGTLSLRKVSSLASEELQSFRNAALGAPGLDKPQQEDLGLPPPPALPPPPAPAPQAPSASVTVSRFSTGTPSNSVNARQALMDAIRSGTGAARLRKVPLLV F4A8D3250294A277 acts_rabit Actin, alpha skeletal muscle ACTA1 ACTA Alpha-actin-1 protein protein rabit Oryctolagus cuniculus (Rabbit) Rabbit MCDEDETTALVCDNGSGLVKAGFAGDDAPRAVFPSIVGRPRHQGVMVGMGQKDSYVGDEAQSKRGILTLKYPIEHGIITNWDDMEKIWHHTFYNELRVAPEEHPTLLTEAPLNPKANREKMTQIMFETFNVPAMYVAIQAVLSLYASGRTTGIVLDSGDGVTHNVPIYEGYALPHAIMRLDLAGRDLTDYLMKILTERGYSFVTTAEREIVRDIKEKLCYVALDFENEMATAASSSSLEKSYELPDGQVITIGNERFRCPETLFQPSFIGMESAGIHETTYNSIMKCDIDIRKDLYANNVMSGGTTMYPGIADRMQKEITALAPSTMKIKIIAPPERKYSVWIGGSILASLSTFQQMWITKQEYDEAGPSIVHRKCF DIP protein P68135 original sequence version: 41 DIP protein P68135 original sequence version: 27 DIP protein P68135 original sequence version: 28 DIP protein P68135 original sequence version: 96 DIP protein P68135 original sequence version: 29 DIP protein P68135 original sequence version: 89 DIP protein P68135 original sequence version: 39 DIP protein P68135 original sequence version: 42 DIP protein P68135 original sequence version: 59 DIP protein P68135 original sequence version: 55 DIP protein P68135 original sequence version: 60 DIP protein P68135 original sequence version: 65 DIP protein P68135 original sequence version: 67 DIP protein P68135 original sequence version: 63 DIP protein P68135 original sequence version: 68 DIP protein P68135 original sequence version: 79 DIP protein P68135 original sequence version: 88 DIP protein P68135 original sequence version: 84 DIP protein P68135 original sequence version: 101 DIP protein P68135 original sequence version: 94 DIP protein P68135 original sequence version: 95 DIP protein P68135 original sequence version: 97 DIP protein P68135 original sequence version: 100 DIP protein P68135 original sequence version: 102 DIP protein P68135 original sequence version: 83 DF2A3A046346A179 pacn1_rat Protein kinase C and casein kinase substrate in neurons protein 1 Pacsin1 Synaptic, dynamin-associated protein I Syndapin-I Dynamin proline-rich domain-interacting protein Syndapin-1 protein protein rat Rattus norvegicus (Rat) Rat MSGPYDEASEEITDSFWEVGNYKRTVKRIDDGHRLCNDLMSCVQERAKIEKAYAQQLTDWAKRWRQLIEKGPQYGSLERAWGAMMTEADKVSELHQEVKNSLLNEDLEKVKNWQKDAYHKQIMGGFKETKEAEDGFRKAQKPWAKKMKELEAAKKAYHLACKEEKLAMTREMNSKTEQSVTPEQQKKLVDKVDKCRQDVQKTQEKYEKVLEDVGKTTPQYMEGMEQVFEQCQQFEEKRLVFLKEVLLDIKRHLNLAENSSYIHVYRELEQAIRGADAQEDLRWFRSTSGPGMPMNWPQFEEWNPDLPHTAAKKEKQPKKAEGAALSNATGAVESTSQAGDRGSVSSYDRGQAYATEWSDDESGNPFGGNEANGGANPFEDDAKGVRVRALYDYDGQEQDELSFKAGDELTKLGEEDEQGWCRGRLDSGQLGLYPANYVEAI DIP protein Q9Z0W5 original sequence version: 106 585B328850572DF8 dbnl_mouse Drebrin-like protein Dbnl Abp1 Sh3p7 SH3 domain-containing protein 7 Actin-binding protein 1 protein protein mouse Mus musculus Mouse MAVNLSRNGPALQEAYVRVVTEKSPTDWALFTYEGNSNDIRVAGTGEGGLEELVEELNSGKVMYAFCRVKDPNSGLPKFVLINWTGEGVNDVRKGACANHVSTMANFLKGAHVTINARAEEDVEPECIMEKVAKASGANYSFHKESTSFQDVGPQAPVGSVYQKTNAISEIKRVGKDNFWAKAEKEEENRRLEEKRRAEEERQRLEEERRERELQEAARREQRYQEQHRSAGAPSPSSRTGEPEQEAVSRTRQEWESAGQQAPHPREIFKQKERAMSTTSVTSSQPGKLRSPFLQKQLTQPETSYGREPTAPVSRPAAGVCEEPAPSTLSSAQTEEEPTYEVPPEQDTLYEEPPLVQQQGAGSEHIDNYMQSQGFSGQGLCARALYDYQAADDTEISFDPENLITGIEVIDEGWWRGYGPDGHFGMFPANYVELIE 85AEF9781C698A3F cobl-pacsin1-1 3 4 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait physical association physical association cobl-pacsin1-2 2 4 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait physical association physical association Figure 1E, Supp Fig S1G Directed yeast two-hybrid analyses of the isolated clone 337 (Y2H 337) as a prey with syndapin I as a bait (E) as well as Y2H 337 as a bait and Abp1 as a prey (F), respectively, cobl-dbnl-1 2 7 unspecified role unspecified role prey prey 4 unspecified role unspecified role bait bait physical association physical association Figure 1F, Supp Fig S1H Directed yeast two-hybrid analyses of the isolated clone 337 (Y2H 337) as a prey with syndapin I as a bait (E) as well as Y2H 337 as a bait and Abp1 as a prey (F), respectively, cobl-acta1-1 1 5 unspecified role unspecified role prey prey region fluorescent dye fluorescent dye label fluorescent dye undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 4 unspecified role unspecified role bait bait wh2-1 region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain wh2-2 region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain wh2-3 region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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 Actin bead assay with Cobl GST-WH2 domains. Figures 2D-2G, 2K-2M Determination of the G-actin dissociation constants for the three WH2 domains by measuring the fluorescence of NBD-labeled actin incubated with different concentrations of the GST-WH2 domain fusion proteins: WH21-1 = 39 nM, WH2-2 = 40 nM , WH2-3 = 432 nM dbnl-acta1-1 1 7 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 5 unspecified role unspecified role prey prey tag fluorescent dye fluorescent dye label fluorescent dye undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association syndapin as well as Abp1 immobilized on beads were still able to induce F-actin halos around the beads (Figures 1B�1D and Figures S1A�S1F Figures 1B-1D, Supp. Figs 1A-1F pacsin1-acta1-1 1 5 unspecified role unspecified role prey prey region fluorescent dye fluorescent dye label fluorescent dye undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 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 ecoli Escherichia coli (strain K12) physical association physical association syndapin as well as Abp1 immobilized on beads were still able to induce F-actin halos around the beads (Figures 1B�1D and Figures S1A�S1F Figures 1B-1D, Supp. Figs 1A-1F