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/ The c-Cbl-associated protein and c-Cbl are two new partners of the SH2-containing inositol polyphosphate 5-phosphatase SHIP2. The c-Cbl-associated protein and c-Cbl are two new partners of the SH2-containing inositol polyphosphate 5-phosphatase SHIP2. Biochem. Biophys. Res. Commun. (0006-291X) 2003 imex curation Vandenbroere I., Paternotte N., Dumont JE., Erneux C., Pirson I. ilpirson@ulb.ac.be 23-Feb-2010: Contacted by JYOTI. Human brain cDNA library jyoti Only protein-protein interactions in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Vandenbroere I., Paternotte N., Dumont JE., Erneux C., Pirson I. ilpirson@ulb.ac.be Accepted 2010-FEB-12 by JYOTI. 23-Feb-2010: Contacted by JYOTI. Human brain cDNA library Biochem. Biophys. Res. Commun. (0006-291X) 2003 jyoti Only protein-protein interactions imex curation The c-Cbl-associated protein and c-Cbl are two new partners of the SH2-containing inositol polyphosphate 5-phosphatase SHIP2. The c-Cbl-associated protein and c-Cbl are two new partners of the SH2-containing inositol polyphosphate 5-phosphatase SHIP2. Biochem. Biophys. Res. Commun. (0006-291X) 2003 imex curation Vandenbroere I., Paternotte N., Dumont JE., Erneux C., Pirson I. ilpirson@ulb.ac.be 23-Feb-2010: Contacted by JYOTI. Human brain cDNA library jyoti Only protein-protein interactions 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 Vandenbroere I., Paternotte N., Dumont JE., Erneux C., Pirson I. ilpirson@ulb.ac.be Accepted 2010-FEB-12 by JYOTI. 23-Feb-2010: Contacted by JYOTI. Human brain cDNA library Biochem. Biophys. Res. Commun. (0006-291X) 2003 jyoti Only protein-protein interactions imex curation The c-Cbl-associated protein and c-Cbl are two new partners of the SH2-containing inositol polyphosphate 5-phosphatase SHIP2. The c-Cbl-associated protein and c-Cbl are two new partners of the SH2-containing inositol polyphosphate 5-phosphatase SHIP2. Biochem. Biophys. Res. Commun. (0006-291X) 2003 imex curation Vandenbroere I., Paternotte N., Dumont JE., Erneux C., Pirson I. ilpirson@ulb.ac.be 23-Feb-2010: Contacted by JYOTI. Human brain cDNA library jyoti Only protein-protein interactions chlae-cos_7 Cercopithecus aethiops simian cells transformed with SV40 Grivet cos_7 cercopithecus aethiops, by transformation with SV40 anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Vandenbroere I., Paternotte N., Dumont JE., Erneux C., Pirson I. ilpirson@ulb.ac.be Accepted 2010-FEB-12 by JYOTI. 23-Feb-2010: Contacted by JYOTI. Human brain cDNA library Biochem. Biophys. Res. Commun. (0006-291X) 2003 jyoti Only protein-protein interactions imex curation The c-Cbl-associated protein and c-Cbl are two new partners of the SH2-containing inositol polyphosphate 5-phosphatase SHIP2. The c-Cbl-associated protein and c-Cbl are two new partners of the SH2-containing inositol polyphosphate 5-phosphatase SHIP2. Biochem. Biophys. Res. Commun. (0006-291X) 2003 imex curation Vandenbroere I., Paternotte N., Dumont JE., Erneux C., Pirson I. ilpirson@ulb.ac.be 23-Feb-2010: Contacted by JYOTI. Human brain cDNA library jyoti Only protein-protein interactions crigr-cho chinese hamster ovary cell line cho chinese hamster ovary cell line Also called LR73 cells. anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Vandenbroere I., Paternotte N., Dumont JE., Erneux C., Pirson I. ilpirson@ulb.ac.be Accepted 2010-FEB-12 by JYOTI. 23-Feb-2010: Contacted by JYOTI. Human brain cDNA library Biochem. Biophys. Res. Commun. (0006-291X) 2003 jyoti Only protein-protein interactions imex curation The c-Cbl-associated protein and c-Cbl are two new partners of the SH2-containing inositol polyphosphate 5-phosphatase SHIP2. The c-Cbl-associated protein and c-Cbl are two new partners of the SH2-containing inositol polyphosphate 5-phosphatase SHIP2. Biochem. Biophys. Res. Commun. (0006-291X) 2003 imex curation Vandenbroere I., Paternotte N., Dumont JE., Erneux C., Pirson I. ilpirson@ulb.ac.be 23-Feb-2010: Contacted by JYOTI. Human brain cDNA library jyoti Only protein-protein interactions 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 Vandenbroere I., Paternotte N., Dumont JE., Erneux C., Pirson I. ilpirson@ulb.ac.be Accepted 2010-FEB-12 by JYOTI. 23-Feb-2010: Contacted by JYOTI. Human brain cDNA library Biochem. Biophys. Res. Commun. (0006-291X) 2003 jyoti Only protein-protein interactions imex curation The c-Cbl-associated protein and c-Cbl are two new partners of the SH2-containing inositol polyphosphate 5-phosphatase SHIP2. The c-Cbl-associated protein and c-Cbl are two new partners of the SH2-containing inositol polyphosphate 5-phosphatase SHIP2. Biochem. Biophys. Res. Commun. (0006-291X) 2003 imex curation Vandenbroere I., Paternotte N., Dumont JE., Erneux C., Pirson I. ilpirson@ulb.ac.be 23-Feb-2010: Contacted by JYOTI. Human brain cDNA library jyoti Only protein-protein interactions crigr-cho chinese hamster ovary cell line cho chinese hamster ovary cell line Also called LR73 cells. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Vandenbroere I., Paternotte N., Dumont JE., Erneux C., Pirson I. ilpirson@ulb.ac.be Accepted 2010-FEB-12 by JYOTI. 23-Feb-2010: Contacted by JYOTI. Human brain cDNA library Biochem. Biophys. Res. Commun. (0006-291X) 2003 jyoti Only protein-protein interactions imex curation ship2_human Phosphatidylinositol 3,4,5-trisphosphate 5-phosphatase 2 INPPL1 SHIP2 Inositol polyphosphate phosphatase-like protein 1 Protein 51C SH2 domain-containing inositol 5'-phosphatase 2 protein protein human Homo sapiens Human MASACGAPGPGGALGSQAPSWYHRDLSRAAAEELLARAGRDGSFLVRDSESVAGAFALCVLYQKHVHTYRILPDGEDFLAVQTSQGVPVRRFQTLGELIGLYAQPNQGLVCALLLPVEGEREPDPPDDRDASDGEDEKPPLPPRSGSTSISAPTGPSSPLPAPETPTAPAAESAPNGLSTVSHDYLKGSYGLDLEAVRGGASHLPHLTRTLATSCRRLHSEVDKVLSGLEILSKVFDQQSSPMVTRLLQQQNLPQTGEQELESLVLKLSVLKDFLSGIQKKALKALQDMSSTAPPAPQPSTRKAKTIPVQAFEVKLDVTLGDLTKIGKSQKFTLSVDVEGGRLVLLRRQRDSQEDWTTFTHDRIRQLIKSQRVQNKLGVVFEKEKDRTQRKDFIFVSARKREAFCQLLQLMKNKHSKQDEPDMISVFIGTWNMGSVPPPKNVTSWFTSKGLGKTLDEVTVTIPHDIYVFGTQENSVGDREWLDLLRGGLKELTDLDYRPIAMQSLWNIKVAVLVKPEHENRISHVSTSSVKTGIANTLGNKGAVGVSFMFNGTSFGFVNCHLTSGNEKTARRNQNYLDILRLLSLGDRQLNAFDISLRFTHLFWFGDLNYRLDMDIQEILNYISRKEFEPLLRVDQLNLEREKHKVFLRFSEEEISFPPTYRYERGSRDTYAWHKQKPTGVRTNVPSWCDRILWKSYPETHIICNSYGCTDDIVTSDHSPVFGTFEVGVTSQFISKKGLSKTSDQAYIEFESIEAIVKTASRTKFFIEFYSTCLEEYKKSFENDAQSSDNINFLKVQWSSRQLPTLKPILADIEYLQDQHLLLTVKSMDGYESYGECVVALKSMIGSTAQQFLTFLSHRGEETGNIRGSMKVRVPTERLGTRERLYEWISIDKDEAGAKSKAPSVSRGSQEPRSGSRKPAFTEASCPLSRLFEEPEKPPPTGRPPAPPRAAPREEPLTPRLKPEGAPEPEGVAAPPPKNSFNNPAYYVLEGVPHQLLPPEPPSPARAPVPSATKNKVAITVPAPQLGHHRHPRVGEGSSSDEESGGTLPPPDFPPPPLPDSAIFLPPSLDPLPGPVVRGRGGAEARGPPPPKAHPRPPLPPGPSPASTFLGEVASGDDRSCSVLQMAKTLSEVDYAPAGPARSALLPGPLELQPPRGLPSDYGRPLSFPPPRIRESIQEDLAEEAPCLQGGRASGLGEAGMSAWLRAIGLERYEEGLVHNGWDDLEFLSDITEEDLEEAGVQDPAHKRLLLDTLQLSK DIP protein O15357 original sequence version: 82 D76B5AA8ACDE8CBA srbs1_human Sorbin and SH3 domain-containing protein 1 KIAA0894 Ponsin c-Cbl-associated protein SH3 domain protein 5 SH3P12 SORBS1 KIAA1296 SH3D5 protein protein human Homo sapiens Human MSSECDGGSKAVMNGLAPGSNGQDKATADPLRARSISAVKIIPVKTVKNASGLVLPTDMDLTKICTGKGAVTLRASSSYRETPSSSPASPQETRQHESKPGLEPEPSSADEWRLSSSADANGNAQPSSLAAKGYRSVHPNLPSDKSQDATSSSAAQPEVIVVPLYLVNTDRGQEGTARPPTPLGPLGCVPTIPATASAASPLTFPTLDDFIPPHLQRWPHHSQPARASGSFAPISQTPPSFSPPPPLVPPAPEDLRRVSEPDLTGAVSSTDSSPLLNEVSSSLIGTDSQAFPSVSKPSSAYPSTTIVNPTIVLLQHNREQQKRLSSLSDPVSERRVGEQDSAPTQEKPTSPGKAIEKRAKDDSRRVVKSTQDLSDVSMDEVGIPLRNTERSKDWYKTMFKQIHKLNRDTPEENPYFPTYKFPELPEIQQTSEEDNPYTPTYQFPASTPSPKSEDDDSDLYSPRYSFSEDTKSPLSVPRSKSEMSYIDGEKVVKRSATLPLPARSSSLKSSSERNDWEPPDKKVDTRKYRAEPKSIYEYQPGKSSVLTNEKMSRDISPEEIDLKNEPWYKFFSELEFGKPPPKKIWDYTPGDCSILPREDRKTNLDKDLSLCQTELEADLEKMETLNKAPSANVPQSSAISPTPEISSETPGYIYSSNFHAVKRESDGAPGDLTSLENERQIYKSVLEGGDIPLQGLSGLKRPSSSASTKDSESPRHFIPADYLESTEEFIRRRHDDKEKLLADQRRLKREQEEADIAARRHTGVIPTHHQFITNERFGDLLNIDDTAKRKSGSEMRPARAKFDFKAQTLKELPLQKGDIVYIYKQIDQNWYEGEHHGRVGIFPRTYIELLPPAEKAQPKKLTPVQVLEYGEAIAKFNFNGDTQVEMSFRKGERITLLRQVDENWYEGRIPGTSRQGIFPITYVDVIKRPLVKNPVDYMDLPFSSSPSRSATASPQFSSHSKLITPAPSSLPHSRRALSPEMHAVTSEWISLTVGVPGRRSLALTPPLPPLPEASIYNTDHLALSPRASPSLSLSLPHLSWSDRPTPRSVASPLALPSPHKTYSLAPTSQASLHMNGDGGVHTPSSGIHQDSFLQLPLGSSDSVISQLSDAFSSQSKRQPWREESGQYERKAERGAGERGPGGPKISKKSCLKPSDVVRCLSTEQRLSDLNTPEESRPGKPLGSAFPGSEAEQTERHRGGEQAGRKAARRGGSQQPQAQQRRVTPDRSQTSQDLFSYQALYSYIPQNDDELELRDGDIVDVMEKCDDGWFVGTSRRTKQFGTFPGNYVKPLYL 70DA4169B6D82F06 a0a3l7ig96_crigr Tyrosine-protein kinase receptor CgPICR_013487 protein protein crigr Cricetulus griseus (Chinese hamster) Cricetulus barabensis griseus Chinese hamster MHLPNVPTVCPSLDIRSEVTELRRLENCSVVEGHLQILLMFAATGEDFRGLSFPRLTQVTDYLLLFRVYGLESLRDLFPNLAVIRGARLFLGYALIIFEMPHLRDVGLPSLGAVLRGAVRVEKNQELCHLSTIDWGLLQPVPGANHIVGNKLGEECADVCPGVLGAAGEPCARTTFSGHTDYRCWTSSHCQRVCPCPHGLACTPRGECCHSECLGGCSQPEDSRACVACRHLYFQGACLRACPPGTYQYESWRCVTAEVCAHLREVPGRTAAFGIYQGSCLAQCPPGFTRNGSSIFCHKCEGLCPKECKVGTKTIDSVQATQDLVGCTHVEGSLILNLRQGYNLEPELQRNLGLVETITGFLKIKHSFALVSLGFFKNLKLIRGDSMVDGNYTLYVLDNQNLQQLGSWVAAGLTIPVGKMYFAFNPRLCLEHIYQLEDVTGTRGRQSKAETDARQEHGQTRTLRFVFNLTEADRILLRWERYEPLEARDLLSFIVYYKESPFQNATEHVGPDACGTQSWNLLDVELPLSRTQEPGVTLAPLKPWTQYAVFVRAITLTTAEDSPHQGAQSPIVYLRTLPAAPTVPQDVISTSNSSSHLLVRWKPPVQRNGNVTYYLVLWQRLAEDGDLYINDYCHRGSLGVPQARGLRLPTSSHDTRFDREDPALEAEPEQGCCPCQHSPPGQALPTLETQEVTFQKKFENFLHHAITIPKSPWKVTSINKRPQRDSERHRREAGPLRQENNSSDFEIQEDKVPRERAVLSGLRHFTEYRIDIHACNHAAHTVGCSAATFVFARTMPHREADDIPGKVAWKAAGRSSVVLHWLEPPDPNGLILKYEIKYRRLGEEATVLCVSRLRYAKVGGVQLALLPPGNYSARVRATSLAGNGSWTDGVAFYITGPEEEDAGRLHILLSVTPVGFMLLIMLAALAFFYSKKRNSTLYTSVNPEYFSASHVYVPDEWEVPREQIAIIRELGQGSFGMVYEGLAQGLEAGEESTPVALKTVNELASARERIEFLKEASVMKAFKCHHVVRLLGVVSQGQPTLVIMELMTRGDLKSHLRSLRPEAENNPGLPQPALSDMIQMAGEIADGMAYLAANKFVHRDLAARNCMVSQDFTVKIGDFGMTRDVYETDYYRKGGKGLLPVRWMAPESLKDGIFTTQSDVWSFGVVLWEIVTLAEQPYQGLSNEQVLKFVMDGGALEELEDCPLQLQELMRLCWQHSPRLRPTFVHILGRIQDELRPSFRLCSFYHSAECQRGQASSLSTETEPDSPPASNGAPDYRPPNGNPGH Similar to P15208 E3FC85140936EA75 a0a061i5t4_crigr E3 ubiquitin-protein ligase CBL H671_4g13114 protein protein crigr Cricetulus griseus (Chinese hamster) Cricetulus barabensis griseus Chinese hamster LQLNSLGPEACRDLRDLLLHNQCQITTLRKFKFNPRLGIDNPVLSLAEDQDQSDPWNLRRPRFCLLSKEEEKSFGFHLQQQLGKADHVVCRVDPGSSAQRHGLREGDRILAVNNKNVEHEDYAMVVRCIRASGPRVLLTVLAQHAHDVVRAQQGSDVFCCPALGPEVRPRLCHIVKDEGGFGFSITHGNRGPFWLVLSSGGAAERAGVPPGARLLEVNGVCVEKFTSNQLSRKVVRLCQNPKLALKNSPPYILDLLPDTYQHLRTVLSRYEGKMETLGENEYFRVFMENLMKKTKQTISLFKEGKERMYEENSQPRRNLTKLSLIFSHMLAELKGIFPSGLFQGDTFRITKADAAEFWRKAFGEKNWNSLAVTHPGYMAFLTYDEVKARLQKFIHKPGSYLFPDGRNQNPDLTGLCEPTPQDHIKVTQEQYELYCEMGSTFQLCKICAENDKDVKIEPCGHLMCTSCLTSWQESEGQGCPFCRCEIKGTEPIVVDPFDPRGSGSLLRQGAEGAPSPNYDDDDDERADDSLFMMKELAGAKAASGSLHKDKPLPIPPTLRDLPPPPPPDRPYSVGTETRPQRRPLPCTPGDCPSRDKLPPVPSSRPGDSWLPRPIPKVPIATPSPNDPWNGRELTNRHSLPFSLPSQMEPRVDVPRLGSTFSLDTSMNMNSSPIAGPENEHPKIKPSSSANAIYSLAARPLPVPKLPPGEQGESEEDAEYMTPTSRPVGAQKPEPKRPLEATQSSRACDCDQQIDSCTYEAMYNIQSQALSVTENSTFGEGNLATAPTSTGPEESENEDDGYDVPKPPVPAVLARRTLSDISNASSSFGWLSLDGDPTTNFNEGSQVPERPPKPFPRRINSERKASSCQQGGATANPSATAPSPQLSSEIERLMSQGYSYQDIQKALVIAHNNIEMAKNILREFVSISSPAHVAT 5E68881894D52209 inppl1-sorbs1-1 5 8 unspecified role unspecified role prey prey region sufficient to bind sufficient binding region sufficient to bind full dna sequence full identification by DNA sequencing full dna sequence certain certain sequence position certain certain certain sequence position certain 7 unspecified role unspecified role bait bait proline rich region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 15 13 physical association physical association Isolation of CAP as SHIP2 partner by yeast two-hybrid screening inppl1-sorbs1-2 2 7 unspecified role unspecified role bait bait proline rich region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 8 unspecified role unspecified role prey prey physical association physical association Isolation of CAP as SHIP2 partner by yeast two-hybrid screening Figs. 1A, 1B inppl1-sorbs1-3 1 7 unspecified role unspecified role prey prey region 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 crigr-cho chinese hamster ovary cell line cho chinese hamster ovary cell line Also called LR73 cells. 8 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 sh3 region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) physical association physical association As shown in Fig. 1C by GST-pulldown, SHIP2 interacted specifically with the GST-SH3C of CAP. Figs. 1C inppl1-sorbs1-4 3 8 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 7 unspecified role unspecified role bait bait region 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 physical association physical association Using anti-CAP immunoblot analysis, we showed (Fig. 2, lower panels) that CAP coimmunoprecipitated with SHIP2. Fig. 2 inppl1-sorbs1-5 4 7 unspecified role unspecified role prey prey region 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 8 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Using anti-CAP immunoblot analysis, we showed (Fig. 2, lower panels) that CAP coimmunoprecipitated with SHIP2. Fig. 3 cbl-ship2 6 7 unspecified role unspecified role prey prey region 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 10 unspecified role unspecified role bait bait physical association physical association Fig. 4 The endogenous SHIP2 was coprecipitated with endogenous c-Cbl as well cbl-sorbs1-1 6 8 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 10 unspecified role unspecified role bait bait physical association physical association Fig. 4 The endogenous SHIP2 was coprecipitated with endogenous c-Cbl as well cbl-sorbs1-2 6 8 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 10 unspecified role unspecified role bait bait 7 unspecified role unspecified role prey prey region 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 physical association physical association Fig. 4 The endogenous SHIP2 was coprecipitated with endogenous c-Cbl as well insr_ship2-1 6 7 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait physical association physical association We also performed immunoprecipitations of the insulin receptor in CHO-IR cells expressing recombinant CAP and SHIP2, and showed the presence of SHIP2 (Fig. 5) in the immunoprecipitate. Fig. 5