MINT MINT, Dpt of Biology, University of Rome Tor Vergata MINT http://mint.bio.uniroma2.it/mint url>http://mint.bio.uniroma2.it/mint/ https://mint.bio.uniroma2.it MINT-[0-9]+ https://mint.bio.uniroma2.it/index.php/results-interactions/?id=${ac} The Molecular INTeraction database (MINT) is a relational database designed to store interactions between biological molecules. http://mint.bio.uniroma2.it http://mint.bio.uniroma2.it/mint disanza-2013-1 CDC42 switches IRSp53 from inhibition of actin growth to elongation by clustering of VASP. in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Disanza A., Bisi S., Winterhoff M., Milanesi F., Ushakov D.S., Kast D., Marighetti P., Romet-Lemonne G., Müller H.M., Nickel W., Linkner J., Waterschoot D., Ampè C., Cortellino S., Palamidessi A., Dominguez R., Carlier M.F., Faix J., Scita G. faix.jan@mh-hannover.de; giorgio.scita@ifom.eu faix.jan@mh-hannover.de,giorgio.scita@ifom.eu EMBO J. (0261-4189) 2013 Only protein-protein interactions imex curation disanza-2013-3 CDC42 switches IRSp53 from inhibition of actin growth to elongation by clustering of VASP. in vitro In vitro dls dynamic light scattering dls predetermined predetermined participant predetermined Disanza A., Bisi S., Winterhoff M., Milanesi F., Ushakov D.S., Kast D., Marighetti P., Romet-Lemonne G., Müller H.M., Nickel W., Linkner J., Waterschoot D., Ampè C., Cortellino S., Palamidessi A., Dominguez R., Carlier M.F., Faix J., Scita G. faix.jan@mh-hannover.de; giorgio.scita@ifom.eu faix.jan@mh-hannover.de,giorgio.scita@ifom.eu EMBO J. (0261-4189) 2013 Only protein-protein interactions imex curation disanza-2013-5 CDC42 switches IRSp53 from inhibition of actin growth to elongation by clustering of VASP. human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Disanza A., Bisi S., Winterhoff M., Milanesi F., Ushakov D.S., Kast D., Marighetti P., Romet-Lemonne G., Müller H.M., Nickel W., Linkner J., Waterschoot D., Ampè C., Cortellino S., Palamidessi A., Dominguez R., Carlier M.F., Faix J., Scita G. faix.jan@mh-hannover.de; giorgio.scita@ifom.eu faix.jan@mh-hannover.de,giorgio.scita@ifom.eu EMBO J. (0261-4189) 2013 Only protein-protein interactions imex curation disanza-2013-4 CDC42 switches IRSp53 from inhibition of actin growth to elongation by clustering of VASP. in vitro In vitro density sedimentation cosedimentation through density gradient density sedimentation predetermined predetermined participant predetermined Disanza A., Bisi S., Winterhoff M., Milanesi F., Ushakov D.S., Kast D., Marighetti P., Romet-Lemonne G., Müller H.M., Nickel W., Linkner J., Waterschoot D., Ampè C., Cortellino S., Palamidessi A., Dominguez R., Carlier M.F., Faix J., Scita G. faix.jan@mh-hannover.de; giorgio.scita@ifom.eu faix.jan@mh-hannover.de,giorgio.scita@ifom.eu EMBO J. (0261-4189) 2013 Only protein-protein interactions imex curation vasp_human Vasodilator-stimulated phosphoprotein VASP protein protein human Homo sapiens Human MSETVICSSRATVMLYDDGNKRWLPAGTGPQAFSRVQIYHNPTANSFRVVGRKMQPDQQVVINCAIVRGVKYNQATPNFHQWRDARQVWGLNFGSKEDAAQFAAGMASALEALEGGGPPPPPALPTWSVPNGPSPEEVEQQKRQQPGPSEHIERRVSNAGGPPAPPAGGPPPPPGPPPPPGPPPPPGLPPSGVPAAAHGAGGGPPPAPPLPAAQGPGGGGAGAPGLAAAIAGAKLRKVSKQEEASGGPTAPKAESGRSGGGGLMEEMNAMLARRRKATQVGEKTPKDESANQEEPEARVPAQSESVRRPWEKNSTTLPRMKSSSSVTTSETQPCTPSSSDYSDLQRVKQELLEEVKKELQKVKEEIIEAFVQELRKRGSP q9uqb8-4 Brain-specific angiogenesis inhibitor 1-associated protein 2 BAIAP2-alpha BAIAP2 Fas ligand-associated factor 3 Insulin receptor substrate p53/p58 Insulin receptor substrate protein of 53 kDa protein protein human Homo sapiens Human MSLSRSEEMHRLTENVYKTIMEQFNPSLRNFIAMGKNYEKALAGVTYAAKGYFDALVKMGELASESQGSKELGDVLFQMAEVHRQIQNQLEEMLKSFHNELLTQLEQKVELDSRYLSAALKKYQTEQRSKGDALDKCQAELKKLRKKSQGSKNPQKYSDKELQYIDAISNKQGELENYVSDGYKTALTEERRRFCFLVEKQCAVAKNSAAYHSKGKELLAQKLPLWQQACADPSKIPERAVQLMQQVASNGATLPSALSASKSNLVISDPIPGAKPLPVPPELAPFVGRMSAQESTPIMNGVTGPDGEDYSPWADRKAAQPKSLSPPQSQSKLSDSYSNTLPVRKSVTPKNSYATTENKTLPRSSSMAAGLERNGRMRVKAIFSHAAGDNSTLLSFKEGDLITLLVPEARDGWHYGESEKTKMRGWFPFSYTRVLDSDGSDRLHMSLQQGKSSSTGNLLDKDDLAIPPPDYGAASRAFPAQTASGFKQRPYSVAVPAFSQGLDDYGARSMSSGSGTLVSTV evl_human Ena/VASP-like protein EVL RNB6 Ena/vasodilator-stimulated phosphoprotein-like protein protein human Homo sapiens Human MSEQSICQARASVMVYDDTSKKWVPIKPGQQGFSRINIYHNTASNTFRVVGVKLQDQQVVINYSIVKGLKYNQATPTFHQWRDARQVYGLNFASKEEATTFSNAMLFALNIMNSQEGGPSSQRQVQNGPSPDEMDIQRRQVMEQHQQQRQESLERRTSATGPILPPGHPSSAASAPVSCSGPPPPPPPPVPPPPTGATPPPPPPLPAGGAQGSSHDESSMSGLAAAIAGAKLRRVQRPEDASGGSSPSGTSKSDANRASSGGGGGGLMEEMNKLLAKRRKAASQSDKPAEKKEDESQMEDPSTSPSPGTRAASQPPNSSEAGRKPWERSNSVEKPVSSILSRTPSVAKSPEAKSPLQSQPHSRMKPAGSVNDMALDAFDLDRMKQEILEEVVRELHKVKEEIIDAIRQELSGISTT cdc42_human Cell division control protein 42 homolog CDC42 G25K GTP-binding protein protein protein human Homo sapiens Human MQTIKCVVVGDGAVGKTCLLISYTTNKFPSEYVPTVFDNYAVTVMIGGEPYTLGLFDTAGQEDYDRLRPLSYPQTDVFLVCFSVVSPSSFENVKEKWVPEITHHCPKTPFLLVGTQIDLRDDPSTIEKLAKNKQKPITPETAEKLARDLKAVKYVECSALTQKGLKNVFDEAILAALEPPEPKKSRRCVLL eps8_human Epidermal growth factor receptor kinase substrate 8 EPS8 protein protein human Homo sapiens Human MNGHISNHPSSFGMYPSQMNGYGSSPTFSQTDREHGSKTSAKALYEQRKNYARDSVSSVSDISQYRVEHLTTFVLDRKDAMITVDDGIRKLKLLDAKGKVWTQDMILQVDDRAVSLIDLESKNELENFPLNTIQHCQAVMHSCSYDSVLALVCKEPTQNKPDLHLFQCDEVKANLISEDIESAISDSKGGKQKRRPDALRMISNADPSIPPPPRAPAPAPPGTVTQVDVRSRVAAWSAWAADQGDFEKPRQYHEQEETPEMMAARIDRDVQILNHILDDIEFFITKLQKAAEAFSELSKRKKNKKGKRKGPGEGVLTLRAKPPPPDEFLDCFQKFKHGFNLLAKLKSHIQNPSAADLVHFLFTPLNMVVQATGGPELASSVLSPLLNKDTIDFLNYTVNGDERQLWMSLGGTWMKARAEWPKEQFIPPYVPRFRNGWEPPMLNFMGATMEQDLYQLAESVANVAEHQRKQEIKRLSTEHSSVSEYHPADGYAFSSNIYTRGSHLDQGEAAVAFKPTSNRHIDRNYEPLKTQPKKYAKSKYDFVARNNSELSVLKDDILEILDDRKQWWKVRNASGDSGFVPNNILDIVRPPESGLGRADPPYTHTIQKQRMEYGPRPADTPPAPSPPPTPAPVPVPLPPSTPAPVPVSKVPANITRQNSSSSDSGGSIVRDSQRHKQLPVDRRKSQMEEVQDELIHRLTIGRSAAQKKFHVPRQNVPVINITYDSTPEDVKTWLQSKGFNPVTVNSLGVLNGAQLFSLNKDELRTVCPEGARVYSQITVQKAALEDSSGSSELQEIMRRRQEKISAAASDSGVESFDEGSSH baiap2-vasp-1 1480896 n/a 1480900 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey purified purified range binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false The range has been inferred by curator. n/a 1480901 western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait purified purified range binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false tag 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 direct interaction direct interaction false false false sf1a sf1d sf1e IRSp53 associates with VASP, as well as EVL, through its SH3 domain (Supplementary Figure S1a–f) that contacts VASP on proline‐rich sites partially overlapping, but distinct from the Profilin binding sites (Supplementary Figure S1c and g). Importantly, the VASP‐ΔPRD mutant, which is unable to bind to IRSp53 (Supplementary Figure S1e), and the IRSp53 W413G mutant, which lacks VASP binding ability (Supplementary Figure S1c), did not form heterocomplexes (Figure 1A, right, and data not shown). In contrast, a VASP mutant lacking the central three GP5 motifs (VASP‐ΔGP5), which retains IRSp53 binding ability, albeit with reduced affinity (Supplementary Figure S1d and f), was still able to hetero‐oligomerize upon addition of IRSp53 (Figure 1A, middle). baiap2-vasp-2 1480896 n/a 1480902 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey purified purified n/a 1480901 predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified range binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false tag 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 direct interaction direct interaction false false false sf1a IRSp53 associates with VASP, as well as EVL, through its SH3 domain (Supplementary Figure S1a–f) that contacts VASP on proline‐rich sites partially overlapping, but distinct from the Profilin binding sites (Supplementary Figure S1c and g). Importantly, the VASP‐ΔPRD mutant, which is unable to bind to IRSp53 (Supplementary Figure S1e), and the IRSp53 W413G mutant, which lacks VASP binding ability (Supplementary Figure S1c), did not form heterocomplexes (Figure 1A, right, and data not shown). In contrast, a VASP mutant lacking the central three GP5 motifs (VASP‐ΔGP5), which retains IRSp53 binding ability, albeit with reduced affinity (Supplementary Figure S1d and f), was still able to hetero‐oligomerize upon addition of IRSp53 (Figure 1A, middle). baiap2-vasp-3 1480896 n/a 1480900 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey purified purified range binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false The range has been inferred by curator. n/a 1480901 western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait purified purified Q9UQB8-4:p.Trp413Gly mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain false tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false direct interaction direct interaction false false false sf1a sf1d sf1e IRSp53 associates with VASP, as well as EVL, through its SH3 domain (Supplementary Figure S1a–f) that contacts VASP on proline‐rich sites partially overlapping, but distinct from the Profilin binding sites (Supplementary Figure S1c and g). Importantly, the VASP‐ΔPRD mutant, which is unable to bind to IRSp53 (Supplementary Figure S1e), and the IRSp53 W413G mutant, which lacks VASP binding ability (Supplementary Figure S1c), did not form heterocomplexes (Figure 1A, right, and data not shown). In contrast, a VASP mutant lacking the central three GP5 motifs (VASP‐ΔGP5), which retains IRSp53 binding ability, albeit with reduced affinity (Supplementary Figure S1d and f), was still able to hetero‐oligomerize upon addition of IRSp53 (Figure 1A, middle). baiap2-vasp-6 1480896 n/a 1480900 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey purified purified n/a 1480901 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey purified purified n/a 1480903 anti tag western anti tag western blot anti tag western unspecified role unspecified role bait bait purified purified tag 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 physical association physical association false false false f3a cdc42-baiap2-6 1480896 n/a 1480903 anti tag western anti tag western blot anti tag western unspecified role unspecified role bait bait purified purified tag 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 1480901 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey purified purified direct interaction direct interaction false false false f3a baiap2-cdc42-2 1480896 n/a 1480903 anti tag western anti tag western blot anti tag western unspecified role unspecified role prey prey purified purified tag 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 1480901 weight by comassie molecular weight estimation by coomasie staining weight by comassie unspecified role unspecified role bait bait purified purified tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1480900 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey purified purified physical association physical association false false false f3b baiap2-cdc42-3 1480896 n/a 1480901 weight by comassie molecular weight estimation by coomasie staining weight by comassie unspecified role unspecified role bait bait purified purified tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false n/a 1480900 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey purified purified direct interaction direct interaction false false false f3b baiap2-vasp-4 1480897 n/a 1480900 predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified range binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain false The range has been inferred by curator. n/a 1480901 predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified direct interaction direct interaction false false false f1a Purified IRSp53 produced a single major species with a hydrodynamic radius of about 7.2 nm (Figure 1A) consistent with its dimeric structure (similar results were obtained by hydrodynamic measurements; Supplementary Figure S2a), while the VASP tetramer gave rise to a single species with a radius of about 14.3 nm (Figure 1A). Notably, the mixture of both proteins led to the formation of large clusters with an average diameter of about 200 nm (Figure 2A, left). Importantly, the VASP‐ΔPRD mutant, which is unable to bind to IRSp53 (Supplementary Figure S1e), and the IRSp53 W413G mutant, which lacks VASP binding ability (Supplementary Figure S1c), did not form heterocomplexes (Figure 1A, right, and data not shown). In contrast, a VASP mutant lacking the central three GP5 motifs (VASP‐ΔGP5), which retains IRSp53 binding ability, albeit with reduced affinity (Supplementary Figure S1d and f), was still able to hetero‐oligomerize upon addition of IRSp53 (Figure 1A, middle). baiap2-vasp-5 1480897 n/a 1480901 predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified Stoichiometry: 2.0 direct interaction direct interaction false false false f1a Purified IRSp53 produced a single major species with a hydrodynamic radius of about 7.2 nm (Figure 1A) consistent with its dimeric structure (similar results were obtained by hydrodynamic measurements; Supplementary Figure S2a), while the VASP tetramer gave rise to a single species with a radius of about 14.3 nm (Figure 1A). Notably, the mixture of both proteins led to the formation of large clusters with an average diameter of about 200 nm (Figure 2A, left). Importantly, the VASP‐ΔPRD mutant, which is unable to bind to IRSp53 (Supplementary Figure S1e), and the IRSp53 W413G mutant, which lacks VASP binding ability (Supplementary Figure S1c), did not form heterocomplexes (Figure 1A, right, and data not shown). In contrast, a VASP mutant lacking the central three GP5 motifs (VASP‐ΔGP5), which retains IRSp53 binding ability, albeit with reduced affinity (Supplementary Figure S1d and f), was still able to hetero‐oligomerize upon addition of IRSp53 (Figure 1A, middle). baiap2-6 1480897 n/a 1480900 predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified Stoichiometry: 4.0 direct interaction direct interaction false false false f1a Purified IRSp53 produced a single major species with a hydrodynamic radius of about 7.2 nm (Figure 1A) consistent with its dimeric structure (similar results were obtained by hydrodynamic measurements; Supplementary Figure S2a), while the VASP tetramer gave rise to a single species with a radius of about 14.3 nm (Figure 1A). Notably, the mixture of both proteins led to the formation of large clusters with an average diameter of about 200 nm (Figure 2A, left). Importantly, the VASP‐ΔPRD mutant, which is unable to bind to IRSp53 (Supplementary Figure S1e), and the IRSp53 W413G mutant, which lacks VASP binding ability (Supplementary Figure S1c), did not form heterocomplexes (Figure 1A, right, and data not shown). In contrast, a VASP mutant lacking the central three GP5 motifs (VASP‐ΔGP5), which retains IRSp53 binding ability, albeit with reduced affinity (Supplementary Figure S1d and f), was still able to hetero‐oligomerize upon addition of IRSp53 (Figure 1A, middle). vasp-baiap2 1480898 n/a 1480900 western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level n/a 1480901 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association false false false The ectopic expression of active CDC42, but not of a dominant‐negative mutant, significantly enhanced the amount of endogeneous IRSp53 co‐immunoprecipitating with VASP (Figure 3C). The CDC42‐dependent increase in the VASP:IRSp53 association was accompanied by a diminution of the EPS8:IRSp53 interaction (Figure 3C). f3c vasp-baiap2-1 1480898 n/a 1480901 western blot western blot Immuno blot Immuno blot unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level n/a 1480904 western blot western blot Immuno blot Immuno blot unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association false false false The ectopic expression of active CDC42, but not of a dominant‐negative mutant, significantly enhanced the amount of endogeneous IRSp53 co‐immunoprecipitating with VASP (Figure 3C). The CDC42‐dependent increase in the VASP:IRSp53 association was accompanied by a diminution of the EPS8:IRSp53 interaction (Figure 3C). f3c baiap2-6 1480899 n/a 1480901 predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified Stoichiometry: 2.0 direct interaction direct interaction false false false sf2a Purified IRSp53 produced a single major species with a hydrodynamic radius of about 7.2 nm (Figure 1A) consistent with its dimeric structure (similar results were obtained by hydrodynamic measurements; Supplementary Figure S2a), while the VASP tetramer gave rise to a single species with a radius of about 14.3 nm (Figure 1A).