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 p85 regulatory subunit controls sequential activation of phosphoinositide 3-kinase by Tyr kinases and Ras. The p85 regulatory subunit controls sequential activation of phosphoinositide 3-kinase by Tyr kinases and Ras. J. Biol. Chem. (0021-9258) 2002 imex curation Jimenez C., Hernandez C., Pimentel B., Carrera AC. Synapse - Interactions of proteins with an established role in the presynapse. Cancer - Interactions investigated in the context of cancer acarrera@cnb.uam.es 16-Dec-2008: Contacted by JYOTI. Only protein-protein interactions in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Jimenez C., Hernandez C., Pimentel B., Carrera AC. Synapse - Interactions of proteins with an established role in the presynapse. Extracts of p65PI3K and p85alpha-expressing CTLL2 cells were incubated with the control column or a Ras-peptide column. After washing, retained material was recovered by boiling beads in SDS sample buffer, resolved by SDS-PAGE, and examined by Western blot. acarrera@cnb.uam.es Accepted 2008-Not a month!-8 by ORCHARD. 16-Dec-2008: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2002 Only protein-protein interactions imex curation The p85 regulatory subunit controls sequential activation of phosphoinositide 3-kinase by Tyr kinases and Ras. The p85 regulatory subunit controls sequential activation of phosphoinositide 3-kinase by Tyr kinases and Ras. J. Biol. Chem. (0021-9258) 2002 imex curation Jimenez C., Hernandez C., Pimentel B., Carrera AC. Synapse - Interactions of proteins with an established role in the presynapse. Cancer - Interactions investigated in the context of cancer acarrera@cnb.uam.es 16-Dec-2008: Contacted by 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 western blot western blot Immuno blot Immuno blot Jimenez C., Hernandez C., Pimentel B., Carrera AC. Synapse - Interactions of proteins with an established role in the presynapse. acarrera@cnb.uam.es Accepted 2008-Not a month!-8 by ORCHARD. 16-Dec-2008: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2002 Only protein-protein interactions imex curation rash_mouse GTPase HRas Hras H-Ras-1 Hras1 Transforming protein p21 c-H-ras p21ras protein protein mouse Mus musculus Mouse MTEYKLVVVGAGGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVVIDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHQYREQIKRVKDSDDVPMVLVGNKCDLAARTVESRQAQDLARSYGIPYIETSAKTRQGVEDAFYTLVREIRQHKLRKLNPPDESGPGCMSCKCVLS DIP protein Q61411 original sequence version: 91 EE6DC2D933E2856A ptn6_human Tyrosine-protein phosphatase non-receptor type 6 PTPN6 HCP Protein-tyrosine phosphatase 1C Hematopoietic cell protein-tyrosine phosphatase SH-PTP1 Protein-tyrosine phosphatase SHP-1 PTP1C protein protein human Homo sapiens Human MVRWFHRDLSGLDAETLLKGRGVHGSFLARPSRKNQGDFSLSVRVGDQVTHIRIQNSGDFYDLYGGEKFATLTELVEYYTQQQGVLQDRDGTIIHLKYPLNCSDPTSERWYHGHMSGGQAETLLQAKGEPWTFLVRESLSQPGDFVLSVLSDQPKAGPGSPLRVTHIKVMCEGGRYTVGGLETFDSLTDLVEHFKKTGIEEASGAFVYLRQPYYATRVNAADIENRVLELNKKQESEDTAKAGFWEEFESLQKQEVKNLHQRLEGQRPENKGKNRYKNILPFDHSRVILQGRDSNIPGSDYINANYIKNQLLGPDENAKTYIASQGCLEATVNDFWQMAWQENSRVIVMTTREVEKGRNKCVPYWPEVGMQRAYGPYSVTNCGEHDTTEYKLRTLQVSPLDNGDLIREIWHYQYLSWPDHGVPSEPGGVLSFLDQINQRQESLPHAGPIIVHCSAGIGRTGTIIVIDMLMENISTKGLDCDIDIQKTIQMVRAQRSGMVQTEAQYKFIYVAIAQFIETTKKKLEVLQSQKGQESEYGNITYPPAMKNAHAKASRTSSKHKEDVYENLHTKNKREEKVKKQRSADKEKSKGSLKRK DIP protein P29350 original sequence version: 183 DIP protein P29350 original sequence version: 139 DIP protein P29350-1 original sequence version: 139 4D7736C21D3542D2 p85a_mouse Phosphatidylinositol 3-kinase regulatory subunit alpha Phosphatidylinositol 3-kinase 85 kDa regulatory subunit alpha Pik3r1 protein protein mouse Mus musculus Mouse MSAEGYQYRALYDYKKEREEDIDLHLGDILTVNKGSLVALGFSDGQEARPEDIGWLNGYNETTGERGDFPGTYVEYIGRKRISPPTPKPRPPRPLPVAPGSSKTEADTEQQALPLPDLAEQFAPPDVAPPLLIKLLEAIEKKGLECSTLYRTQSSSNPAELRQLLDCDAASVDLEMIDVHVLADAFKRYLADLPNPVIPVAVYNEMMSLAQELQSPEDCIQLLKKLIRLPNIPHQCWLTLQYLLKHFFKLSQASSKNLLNARVLSEIFSPVLFRFPAASSDNTEHLIKAIEILISTEWNERQPAPALPPKPPKPTTVANNSMNNNMSLQDAEWYWGDISREEVNEKLRDTADGTFLVRDASTKMHGDYTLTLRKGGNNKLIKIFHRDGKYGFSDPLTFNSVVELINHYRNESLAQYNPKLDVKLLYPVSKYQQDQVVKEDNIEAVGKKLHEYNTQFQEKSREYDRLYEEYTRTSQEIQMKRTAIEAFNETIKIFEEQCQTQERYSKEYIEKFKREGNEKEIQRIMHNHDKLKSRISEIIDSRRRLEEDLKKQAAEYREIDKRMNSIKPDLIQLRKTRDQYLMWLTQKGVRQKKLNEWLGNENTEDQYSLVEDDEDLPHHDEKTWNVGSSNRNKAENLLRGKRDGTFLVRESSKQGCYACSVVVDGEVKHCVINKTATGYGFAEPYNLYSSLKELVLHYQHTSLVQHNDSLNVTLAYPVYAQQRR DIP protein P26450 original sequence version: 149 DIP protein P26450 original sequence version: 142 DIP protein P26450 original sequence version: 140 9975D7AD8BABBA9C pik3r1-shp1 2 5 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 4 unspecified role unspecified role prey prey physical association physical association 3 when p65PI3K and p85 regulatory subunits were immunopurified with anti-HA Ab, both subunits recruited similar amounts of SHP-1 When p85alpha and the p65 isoform were immunopurified with anti-HA antibodies, both subunits recruited similar amounts of SHP-1, suggesting that p85 binding to SHP-1 is not mediated exclusively by the C-SH2 domain. A p65 isoform of Pik3r1 was also shown to interact with PTPN6 but this isoform could not be matched to any sequence in the UniProt database at the time of curation. hras-pik3r1 1 3 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain chemical synthesis Chemical synthesis (Chemical synthesis) 5 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 mouse-ctll_2 Mus musculus CTLL-2 C57b1/6 T cell lymphoblasts ctll_2 CTLL-2 mouse C57b1/6 T cell lymphoblasts physical association physical association 5 extracts of p65PI3K- and p85-expressing CTLL2 cells were prepared and incubated with the Ras peptide-specific column or a control column. p65PI3K and p85 were both retained in the Ras column, with slightly greater retention of p85 than of p65PI3K A p65 isoform of Pik3r1 (P26450) was also shown to bind but this could not be matched to any sequence in UniProt at the time of annotation.