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/ Mouse profilin 2 regulates endocytosis and competes with SH3 ligand binding to dynamin 1. Mouse profilin 2 regulates endocytosis and competes with SH3 ligand binding to dynamin 1. J. Biol. Chem. (0021-9258) 2006 imex curation Gareus R., Di Nardo A., Rybin V., Witke W. BioCreative - Critical Assessment of Information Extraction systems in Biology Synapse - Interactions of proteins with an established role in the presynapse. Fig 3 ELISA data could not be added since protein species could not be traced. Author contacted - no repsonse. witke@embl-monterotondo.it 24-JAN-2007: Contacted by JYOTI. dave Only protein-protein interactions in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Gareus R., Di Nardo A., Rybin V., Witke W. BioCreative - Critical Assessment of Information Extraction systems in Biology Fig 3 ELISA data could not be added since protein species could not be traced. Author contacted - no repsonse. Bait linked to beads - linkage methodology and composition of beads not stated in publication. witke@embl-monterotondo.it Accepted 2006-SEP-14 by DAVET. 24-JAN-2007: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2006 dave Only protein-protein interactions imex curation Mouse profilin 2 regulates endocytosis and competes with SH3 ligand binding to dynamin 1. Mouse profilin 2 regulates endocytosis and competes with SH3 ligand binding to dynamin 1. J. Biol. Chem. (0021-9258) 2006 imex curation Gareus R., Di Nardo A., Rybin V., Witke W. BioCreative - Critical Assessment of Information Extraction systems in Biology Synapse - Interactions of proteins with an established role in the presynapse. Fig 3 ELISA data could not be added since protein species could not be traced. Author contacted - no repsonse. witke@embl-monterotondo.it 24-JAN-2007: Contacted by JYOTI. dave Only protein-protein interactions in vitro In vitro solution sedimentati cosedimentation in solution solution sedimentati predetermined predetermined participant predetermined Gareus R., Di Nardo A., Rybin V., Witke W. BioCreative - Critical Assessment of Information Extraction systems in Biology Fig 3 ELISA data could not be added since protein species could not be traced. Author contacted - no repsonse. Experiments were done at 20 �C using a two-channel charcoal centerpiece (3-mm path length) at a speed of 55,000 rpm in a Beckman Optima XL-A centrifuge fitted with a four-hole AN-60 rotor. witke@embl-monterotondo.it Accepted 2006-SEP-14 by DAVET. 24-JAN-2007: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2006 dave Only protein-protein interactions imex curation Mouse profilin 2 regulates endocytosis and competes with SH3 ligand binding to dynamin 1. Mouse profilin 2 regulates endocytosis and competes with SH3 ligand binding to dynamin 1. J. Biol. Chem. (0021-9258) 2006 imex curation Gareus R., Di Nardo A., Rybin V., Witke W. BioCreative - Critical Assessment of Information Extraction systems in Biology Synapse - Interactions of proteins with an established role in the presynapse. Fig 3 ELISA data could not be added since protein species could not be traced. Author contacted - no repsonse. witke@embl-monterotondo.it 24-JAN-2007: Contacted by JYOTI. dave Only protein-protein interactions mouse-brain Mus musculus brain brain Brain [cerebrum] anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Gareus R., Di Nardo A., Rybin V., Witke W. BioCreative - Critical Assessment of Information Extraction systems in Biology Fig 3 ELISA data could not be added since protein species could not be traced. Author contacted - no repsonse. witke@embl-monterotondo.it Accepted 2006-SEP-14 by DAVET. J. Biol. Chem. (0021-9258) 2006 dave Only protein-protein interactions imex curation Mouse profilin 2 regulates endocytosis and competes with SH3 ligand binding to dynamin 1. Mouse profilin 2 regulates endocytosis and competes with SH3 ligand binding to dynamin 1. J. Biol. Chem. (0021-9258) 2006 imex curation Gareus R., Di Nardo A., Rybin V., Witke W. BioCreative - Critical Assessment of Information Extraction systems in Biology Synapse - Interactions of proteins with an established role in the presynapse. Fig 3 ELISA data could not be added since protein species could not be traced. Author contacted - no repsonse. witke@embl-monterotondo.it 24-JAN-2007: Contacted by JYOTI. dave Only protein-protein interactions in vitro In vitro peptide array peptide array antibody detection identification by antibody antibody detection Gareus R., Di Nardo A., Rybin V., Witke W. BioCreative - Critical Assessment of Information Extraction systems in Biology Fig 3 ELISA data could not be added since protein species could not be traced. Author contacted - no repsonse. 32 12-mer peptides covering the entire PRD of dynamin 1 were synthesized on a SPOT membrane witke@embl-monterotondo.it Accepted 2006-SEP-14 by DAVET. 24-JAN-2007: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2006 dave Only protein-protein interactions imex curation prof2_mouse Profilin-2 Profilin II Pfn2 protein protein mouse Mus musculus Mouse MAGWQSYVDNLMCDGCCQEAAIVGYCDAKYVWAATAGGVFQSITPVEIDMIVGKDREGFFTNGLTLGAKKCSVIRDSLYVDGDCTMDIRTKSQGGEPTYNVAVGRAGRVLVFVMGKEGVHGGGLNKKAYSMAKYLRDSGF E780BF62E9EC8476 q9jjv2-1 Profilin-2 Pfn2 Profilin II IIa protein protein mouse Mus musculus Mouse MAGWQSYVDNLMCDGCCQEAAIVGYCDAKYVWAATAGGVFQSITPVEIDMIVGKDREGFFTNGLTLGAKKCSVIRDSLYVDGDCTMDIRTKSQGGEPTYNVAVGRAGRVLVFVMGKEGVHGGGLNKKAYSMAKYLRDSGF E780BF62E9EC8476 p39053-4 Dynamin-1 BraDnm2 Dnm1 Dnm Kiaa4093 protein protein mouse Mus musculus Mouse MGNRGMEDLIPLVNRLQDAFSAIGQNADLDLPQIAVVGGQSAGKSSVLENFVGRDFLPRGSGIVTRRPLVLQLVNSTTEYAEFLHCKGKKFTDFEEVRLEIEAETDRVTGTNKGISPVPINLRVYSPHVLNLTLVDLPGMTKVPVGDQPPDIEFQIRDMLMQFVTKENCLILAVSPANSDLANSDALKIAKEVDPQGQRTIGVITKLDLMDEGTDARDVLENKLLPLRRGYIGVVNRSQKDIDGKKDITAALAAERKFFLSHPSYRHLADRMGTPYLQKVLNQQLTNHIRDTLPGLRNKLQSQLLSIEKEVDEYKNFRPDDPARKTKALLQMVQQFAVDFEKRIEGSGDQIDTYELSGGARINRIFHERFPFELVKMEFDEKELRREISYAIKNIHGIRTGLFTPDMAFETIVKKQVKKIREPCLKCVDMVISELISTVRQCTKKLQQYPRLREEMERIVTTHIREREGRTKEQVMLLIDIELAYMNTNHEDFIGFANAQQRSNQMNKKKTSGNQDEILVIRKGWLTINNIGIMKGGSKEYWFVLTAENLSWYKDDEEKEKKYMLSVDNLKLRDVEKGFMSSKHIFALFNTEQRNVYKDYRQLELACETQEEVDSWKASFLRAGVYPERVGDKEKASETEENGSDSFMHSMDPQLERQVETIRNLVDSYMAIVNKTVRDLMPKTIMHLMINNTKEFIFSELLANLYSCGDQNTLMEESAEQAQRRDEMLRMYHALKEALSIIGDINTTTVSTPMPPPVDDSWLQVQSVPAGRRSPTSSPTPQRRAPAVPPARPGSRGPAPGPPPAGSALGGAPPVPSRPGASPDPFGPPPQVPSRPNRAPPGVPSRSGQASPSRPESPRPPFDL Contains a phosphoserine at position 857. Contains a phosphoserine at position 851. 4E27474E413CCEE7 dyn1_mouse Dynamin-1 Dnm1 Dnm Kiaa4093 protein protein mouse Mus musculus Mouse MGNRGMEDLIPLVNRLQDAFSAIGQNADLDLPQIAVVGGQSAGKSSVLENFVGRDFLPRGSGIVTRRPLVLQLVNSTTEYAEFLHCKGKKFTDFEEVRLEIEAETDRVTGTNKGISPVPINLRVYSPHVLNLTLVDLPGMTKVPVGDQPPDIEFQIRDMLMQFVTKENCLILAVSPANSDLANSDALKIAKEVDPQGQRTIGVITKLDLMDEGTDARDVLENKLLPLRRGYIGVVNRSQKDIDGKKDITAALAAERKFFLSHPSYRHLADRMGTPYLQKVLNQQLTNHIRDTLPGLRNKLQSQLLSIEKEVDEYKNFRPDDPARKTKALLQMVQQFAVDFEKRIEGSGDQIDTYELSGGARINRIFHERFPFELVKMEFDEKELRREISYAIKNIHGIRTGLFTPDMAFETIVKKQVKKIREPCLKCVDMVISELISTVRQCTKKLQQYPRLREEMERIVTTHIREREGRTKEQVMLLIDIELAYMNTNHEDFIGFANAQQRSNQMNKKKTSGNQDEILVIRKGWLTINNIGIMKGGSKEYWFVLTAENLSWYKDDEEKEKKYMLSVDNLKLRDVEKGFMSSKHIFALFNTEQRNVYKDYRQLELACETQEEVDSWKASFLRAGVYPERVGDKEKASETEENGSDSFMHSMDPQLERQVETIRNLVDSYMAIVNKTVRDLMPKTIMHLMINNTKEFIFSELLANLYSCGDQNTLMEESAEQAQRRDEMLRMYHALKEALSIIGDINTTTVSTPMPPPVDDSWLQVQSVPAGRRSPTSSPTPQRRAPAVPPARPGSRGPAPGPPPAGSALGGAPPVPSRPGASPDPFGPPPQVPSRPNRAPPGVPSLGAWRLNSPQGKHENRAGKARL F48EC3DF5F39A08B sh3g2_mouse Endophilin-A1 Sh3gl2 Sh3d2a Endophilin-1 SH3 domain-containing GRB2-like protein 2 SH3 domain protein 2A Een1 SH3p4 protein protein mouse Mus musculus Mouse MSVAGLKKQFHKATQKVSEKVGGAEGTKLDDDFKEMERKVDVTSRAVMEIMTKTIEYLQPNPASRAKLSMINTMSKIRGQEKGPGYPQAEALLAEAMLKFGRELGDDCNFGPALGEVGEAMRELSEVKDSLDMEVKQNFIDPLQNLHDKDLREIQHHLKKLEGRRLDFDYKKKRQGKIPDEELRQALEKFDESKEIAESSMFNLLEMDIEQVSQLSALVQAQLEYHKQAVQILQQVTVRLEERIRQASSQPRREYQPKPRMSLEFATGDSTQPNGGLSHTGTPKPPGVQMDQPCCRALYDFEPENEGELGFKEGDIITLTNQIDENWYEGMLHGQSGFFPINYVEILVALPH DIP protein Q62420 original sequence version: 77 8C2B6F0FB245DCD3 vasp_mouse Vasodilator-stimulated phosphoprotein Vasp protein protein mouse Mus musculus Mouse MSETVICSSRATVMLYDDSNKRWLPAGTGPQAFSRVQIYHNPTANSFRVVGRKMQPDQQVVINCAIIRGVKYNQATPIFHQWRDARQVWGLNFGSKEDAIQFATGMANALEALEGGGPPPAPAPPAWSAQNGPSPEELEQQKRQPEHMERRVSNAGGPPAPPAGGPPPPPGPPPPPGPPPPPGLPSSGVSGAGHGAGAAPPPAPPLPTAQGPNSGGSGAPGLAAAIAGAKLRKVSKQEEASGGPLAPKAENSRSTGGGLMEEMNAMLARRRKATQVGEKPPKDESASEESEARLPAQSEPVRRPWEKNSTTLPRMKSSSSVTTSEAHPSTPCSSDDSDLERVKQELLEEVRKELQKMKEEIIEVFVQELRKRGSP DIP protein P70460 original sequence version: 128 19369286CF4276C7 dnm1-pfn2-1 3 8 unspecified role unspecified role prey prey 5 unspecified role unspecified role bait bait physical association physical association As shown in Fig. 1A, dynamin 1 can be co-immunoprecipitated with profilin 2 from brain lysates using a profilin 2-specific antibody. 1A dnm1-pfn2-2 1 8 unspecified role unspecified role prey prey proline rich region required to bind necessary binding region required to bind deletion analysis deletion analysis range range range range spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection 6 unspecified role unspecified role bait bait Q9JJV2-1:p.Ser138Asp mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain S D Wed May 02 19:16:17 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser138asp' Tue Jun 30 19:33:13 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser138Asp' ecoli Escherichia coli (strain K12) direct interaction direct interaction To test for direct binding, we expressed and purified recombinant dynamin 1 and performed binding studies using beads loaded with profilin 1 and profilin 2 (Fig. 1B). 1B, 1C, 1D, 1E Publication describes an isoform of Dynamin-1, Dynamin 1aa, as the recombinant protein but this description cannot be matched to any of the isoforms within UniProtKB. By poly-L-proline, suggesting that the poly-L-proline binding site of profilin is required for dynamin 1 binding. A second Ser-133->Tyr Profilin-2 mutant is described in the paper but the UniProtKB entry already shows a tyrosine in this position. dnm1-pfn2-3 4 7 unspecified role unspecified role bait bait region (strong binding sites) sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain region (weak binding sites) sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain in vitro In vitro 6 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) direct interaction direct interaction To identify the exact binding sites of profilin 2 within the PRD and to compare these with the docking sites of other known dynamin 1 ligands we made use of a SPOT peptide array covering the entire PRD. 1F, Table 1 dnm1-sh3gl2-1 4 7 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 certain certain sequence position certain certain certain sequence position certain in vitro In vitro 9 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) direct interaction direct interaction To identify the exact binding sites of profilin 2 within the PRD and to compare these with the docking sites of other known dynamin 1 ligands we made use of a SPOT peptide array covering the entire PRD. 1F, Table 1 dnm1-dnm1-1 2 8 unspecified role unspecified role neutral component neutral component 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 spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection direct interaction direct interaction 1G Although we have identified several potential profilin 2 binding sites along the PRD, the question remains whether all these sites are occupied in the profilin 2-dynamin 1 complex. To determine the stoichiometry of the complex in vitro we performed velocity sedimentation using an analytical ultracentrifuge. dnm1-dnm1-2 2 8 unspecified role unspecified role neutral component neutral component 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 spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection 6 unspecified role unspecified role neutral component neutral component ecoli Escherichia coli (strain K12) direct interaction direct interaction 1G Although we have identified several potential profilin 2 binding sites along the PRD, the question remains whether all these sites are occupied in the profilin 2-dynamin 1 complex. To determine the stoichiometry of the complex in vitro we performed velocity sedimentation using an analytical ultracentrifuge. vasp-pfn2-1 4 10 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 in vitro In vitro 6 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) direct interaction direct interaction To identify the exact binding sites of profilin 2 within the PRD and to compare these with the docking sites of other known dynamin 1 ligands we made use of a SPOT peptide array covering the entire PRD. 1F, Table 1 Positive control