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 p85alpha subunit of phosphatidylinositol 3'-kinase binds to and stimulates the GTPase activity of Rab proteins. The p85alpha subunit of phosphatidylinositol 3'-kinase binds to and stimulates the GTPase activity of Rab proteins. J. Biol. Chem. (0021-9258) 2004 imex curation Chamberlain MD., Berry TR., Pastor MC., Anderson DH. Synapse - Interactions of proteins with an established role in the presynapse. Cancer - Interactions investigated in the context of cancer Cells were stimulated with PDGF BB for various times before immunoprecipitation was carried out. danderson@scf.sk.ca The antibody used for p85 was specific to the alpha isoform of p85, as described on provider's website (Upstate Biotechnology - now Millipore - cat. no. 05-217). The antibody used for PDGFR was specific for the beta isoform, as decribed on the provider's website (Santa Cruz Biotechnology , cat. no. sc-432). orchard Only protein-protein interactions chlae-cos_1 Chlorocebus aethiops COS-1 cells Grivet cos_1 CV-1 Simian cells (cercopithecus aethiops) transformed by SV40 anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Chamberlain MD., Berry TR., Pastor MC., Anderson DH. Synapse - Interactions of proteins with an established role in the presynapse. Cells were stimulated with PDGF BB for various times before immunoprecipitation was carried out. danderson@scf.sk.ca Accepted 2007-DEC-28 by ORCHARD. The antibody used for p85 was specific to the alpha isoform of p85, as described on provider's website (Upstate Biotechnology - now Millipore - cat. no. 05-217). The antibody used for PDGFR was specific for the beta isoform, as decribed on the provider's website (Santa Cruz Biotechnology , cat. no. sc-432). J. Biol. Chem. (0021-9258) 2004 orchard Only protein-protein interactions imex curation The p85alpha subunit of phosphatidylinositol 3'-kinase binds to and stimulates the GTPase activity of Rab proteins. The p85alpha subunit of phosphatidylinositol 3'-kinase binds to and stimulates the GTPase activity of Rab proteins. J. Biol. Chem. (0021-9258) 2004 imex curation Chamberlain MD., Berry TR., Pastor MC., Anderson DH. Synapse - Interactions of proteins with an established role in the presynapse. Cancer - Interactions investigated in the context of cancer Cells were stimulated with PDGF BB for various times before immunoprecipitation was carried out. danderson@scf.sk.ca The antibody used for p85 was specific to the alpha isoform of p85, as described on provider's website (Upstate Biotechnology - now Millipore - cat. no. 05-217). The antibody used for PDGFR was specific for the beta isoform, as decribed on the provider's website (Santa Cruz Biotechnology , cat. no. sc-432). orchard Only protein-protein interactions mouse-nih_3t3 Mus musculus Swiss NIH 3t3 embryo fibroblast cells nih_3t3 Mouse Swiss NIH embryo contact inhibited fibroblast cells anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Chamberlain MD., Berry TR., Pastor MC., Anderson DH. Synapse - Interactions of proteins with an established role in the presynapse. Cells were stimulated with PDGF BB for various times before immunoprecipitation was carried out. danderson@scf.sk.ca Accepted 2007-DEC-28 by ORCHARD. The antibody used for p85 was specific to the alpha isoform of p85, as described on provider's website (Upstate Biotechnology - now Millipore - cat. no. 05-217). The antibody used for PDGFR was specific for the beta isoform, as decribed on the provider's website (Santa Cruz Biotechnology , cat. no. sc-432). J. Biol. Chem. (0021-9258) 2004 orchard Only protein-protein interactions imex curation The p85alpha subunit of phosphatidylinositol 3'-kinase binds to and stimulates the GTPase activity of Rab proteins. The p85alpha subunit of phosphatidylinositol 3'-kinase binds to and stimulates the GTPase activity of Rab proteins. J. Biol. Chem. (0021-9258) 2004 imex curation Chamberlain MD., Berry TR., Pastor MC., Anderson DH. Synapse - Interactions of proteins with an established role in the presynapse. Cancer - Interactions investigated in the context of cancer Cells were stimulated with PDGF BB for various times before immunoprecipitation was carried out. danderson@scf.sk.ca The antibody used for p85 was specific to the alpha isoform of p85, as described on provider's website (Upstate Biotechnology - now Millipore - cat. no. 05-217). The antibody used for PDGFR was specific for the beta isoform, as decribed on the provider's website (Santa Cruz Biotechnology , cat. no. sc-432). orchard Only protein-protein interactions in vitro In vitro elisa enzyme linked immunosorbent assay ELISA ELISA elisa antibody detection identification by antibody antibody detection Chamberlain MD., Berry TR., Pastor MC., Anderson DH. Synapse - Interactions of proteins with an established role in the presynapse. Cells were stimulated with PDGF BB for various times before immunoprecipitation was carried out. danderson@scf.sk.ca Accepted 2007-DEC-28 by ORCHARD. The antibody used for p85 was specific to the alpha isoform of p85, as described on provider's website (Upstate Biotechnology - now Millipore - cat. no. 05-217). The antibody used for PDGFR was specific for the beta isoform, as decribed on the provider's website (Santa Cruz Biotechnology , cat. no. sc-432). J. Biol. Chem. (0021-9258) 2004 orchard Only protein-protein interactions imex curation The p85alpha subunit of phosphatidylinositol 3'-kinase binds to and stimulates the GTPase activity of Rab proteins. The p85alpha subunit of phosphatidylinositol 3'-kinase binds to and stimulates the GTPase activity of Rab proteins. J. Biol. Chem. (0021-9258) 2004 imex curation Chamberlain MD., Berry TR., Pastor MC., Anderson DH. Synapse - Interactions of proteins with an established role in the presynapse. Cancer - Interactions investigated in the context of cancer Cells were stimulated with PDGF BB for various times before immunoprecipitation was carried out. danderson@scf.sk.ca The antibody used for p85 was specific to the alpha isoform of p85, as described on provider's website (Upstate Biotechnology - now Millipore - cat. no. 05-217). The antibody used for PDGFR was specific for the beta isoform, as decribed on the provider's website (Santa Cruz Biotechnology , cat. no. sc-432). orchard Only protein-protein interactions mouse-nih_3t3 Mus musculus Swiss NIH 3t3 embryo fibroblast cells nih_3t3 Mouse Swiss NIH embryo contact inhibited fibroblast cells anti bait coip anti bait coimmunoprecipitation anti bait coip anti tag western anti tag western blot anti tag western Chamberlain MD., Berry TR., Pastor MC., Anderson DH. Synapse - Interactions of proteins with an established role in the presynapse. Cells were stimulated with PDGF BB for various times before immunoprecipitation was carried out. danderson@scf.sk.ca Accepted 2007-DEC-28 by ORCHARD. The antibody used for p85 was specific to the alpha isoform of p85, as described on provider's website (Upstate Biotechnology - now Millipore - cat. no. 05-217). The antibody used for PDGFR was specific for the beta isoform, as decribed on the provider's website (Santa Cruz Biotechnology , cat. no. sc-432). J. Biol. Chem. (0021-9258) 2004 orchard Only protein-protein interactions imex curation The p85alpha subunit of phosphatidylinositol 3'-kinase binds to and stimulates the GTPase activity of Rab proteins. The p85alpha subunit of phosphatidylinositol 3'-kinase binds to and stimulates the GTPase activity of Rab proteins. J. Biol. Chem. (0021-9258) 2004 imex curation Chamberlain MD., Berry TR., Pastor MC., Anderson DH. Synapse - Interactions of proteins with an established role in the presynapse. Cancer - Interactions investigated in the context of cancer Cells were stimulated with PDGF BB for various times before immunoprecipitation was carried out. danderson@scf.sk.ca The antibody used for p85 was specific to the alpha isoform of p85, as described on provider's website (Upstate Biotechnology - now Millipore - cat. no. 05-217). The antibody used for PDGFR was specific for the beta isoform, as decribed on the provider's website (Santa Cruz Biotechnology , cat. no. sc-432). orchard Only protein-protein interactions in vitro In vitro gtpase assay gtpase assay GTPase gtp hydrolisis gtp hydrolisis GTPase weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Chamberlain MD., Berry TR., Pastor MC., Anderson DH. Synapse - Interactions of proteins with an established role in the presynapse. Cells were stimulated with PDGF BB for various times before immunoprecipitation was carried out. danderson@scf.sk.ca Accepted 2007-DEC-28 by ORCHARD. The antibody used for p85 was specific to the alpha isoform of p85, as described on provider's website (Upstate Biotechnology - now Millipore - cat. no. 05-217). The antibody used for PDGFR was specific for the beta isoform, as decribed on the provider's website (Santa Cruz Biotechnology , cat. no. sc-432). J. Biol. Chem. (0021-9258) 2004 orchard Only protein-protein interactions imex curation The p85alpha subunit of phosphatidylinositol 3'-kinase binds to and stimulates the GTPase activity of Rab proteins. The p85alpha subunit of phosphatidylinositol 3'-kinase binds to and stimulates the GTPase activity of Rab proteins. J. Biol. Chem. (0021-9258) 2004 imex curation Chamberlain MD., Berry TR., Pastor MC., Anderson DH. Synapse - Interactions of proteins with an established role in the presynapse. Cancer - Interactions investigated in the context of cancer Cells were stimulated with PDGF BB for various times before immunoprecipitation was carried out. danderson@scf.sk.ca The antibody used for p85 was specific to the alpha isoform of p85, as described on provider's website (Upstate Biotechnology - now Millipore - cat. no. 05-217). The antibody used for PDGFR was specific for the beta isoform, as decribed on the provider's website (Santa Cruz Biotechnology , cat. no. sc-432). orchard Only protein-protein interactions in vitro In vitro pull down pull down sequence tag sequence tag identification sequence tag Chamberlain MD., Berry TR., Pastor MC., Anderson DH. Synapse - Interactions of proteins with an established role in the presynapse. Cells were stimulated with PDGF BB for various times before immunoprecipitation was carried out. danderson@scf.sk.ca Accepted 2007-DEC-28 by ORCHARD. The antibody used for p85 was specific to the alpha isoform of p85, as described on provider's website (Upstate Biotechnology - now Millipore - cat. no. 05-217). The antibody used for PDGFR was specific for the beta isoform, as decribed on the provider's website (Santa Cruz Biotechnology , cat. no. sc-432). J. Biol. Chem. (0021-9258) 2004 orchard Only protein-protein interactions imex curation rab5a_human Ras-related protein Rab-5A RAB5A RAB5 protein protein human Homo sapiens Human MASRGATRPNGPNTGNKICQFKLVLLGESAVGKSSLVLRFVKGQFHEFQESTIGAAFLTQTVCLDDTTVKFEIWDTAGQERYHSLAPMYYRGAQAAIVVYDITNEESFARAKNWVKELQRQASPNIVIALSGNKADLANKRAVDFQEAQSYADDNSLLFMETSAKTSMNVNEIFMAIAKKLPKNEPQNPGANSARGRGVDLTEPTQPTRNQCCSN DIP protein P20339 original sequence version: 100 EC03DDF96BBEF821 pk3ca_mouse Phosphatidylinositol 4,5-bisphosphate 3-kinase catalytic subunit alpha isoform Phosphoinositide-3-kinase catalytic alpha polypeptide Serine/threonine protein kinase PIK3CA Pik3ca Phosphatidylinositol 4,5-bisphosphate 3-kinase 110 kDa catalytic subunit alpha protein protein mouse Mus musculus Mouse MPPRPSSGELWGIHLMPPRILVECLLPNGMIVTLECLREATLVTIKHELFREARKYPLHQLLQDETSYIFVSVTQEAEREEFFDETRRLCDLRLFQPFLKVIEPVGNREEKILNREIGFVIGMPVCEFDMVKDPEVQDFRRNILNVCKEAVDLRDLNSPHSRAMYVYPPNVESSPELPKHIYNKLDKGQIIVVIWVIVSPNNDKQKYTLKINHDCVPEQVIAEAIRKKTRSMLLSSEQLKLCVLEYQGKYILKVCGCDEYFLEKYPLSQYKYIRSCIMLGRMPNLMLMAKESLYSQLPIDSFTMPSYSRRISTATPYMNGETSTKSLWVINSALRIKILCATYVNVNIRDIDKIYVRTGIYHGGEPLCDNVNTQRVPCSNPRWNEWLNYDIYIPDLPRAARLCLSICSVKGRKGAKEEHCPLAWGNINLFDYTDTLVSGKMALNLWPVPHGLEDLLNPIGVTGSNPNKETPCLELEFDWFSSVVKFPDMSVIEEHANWSVSREAGFSYSHTGLSNRLARDNELRENDKEQLRALCTRDPLSEITEQEKDFLWSHRHYCVTIPEILPKLLLSVKWNSRDEVAQMYCLVKDWPPIKPEQAMELLDCNYPDPMVRSFAVRCLEKYLTDDKLSQYLIQLVQVLKYEQYLDNLLVRFLLKKALTNQRIGHFFFWHLKSEMHNKTVSQRFGLLLESYCRACGMYLKHLNRQVEAMEKLINLTDILKQEKKDETQKVQMKFLVEQMRQPDFMDALQGFLSPLNPAHQLGNLRLEECRIMSSAKRPLWLNWENPDIMSELLFQNNEIIFKNGDDLRQDMLTLQIIRIMENIWQNQGLDLRMLPYGCLSIGDCVGLIEVVRNSHTIMQIQCKGGLKGALQFNSHTLHQWLKDKNKGEIYDAAIDLFTRSCAGYCVATFILGIGDRHNSNIMVKDDGQLFHIDFGHFLDHKKKKFGYKRERVPFVLTQDFLIVISKGAQEYTKTREFERFQEMCYKAYLAIRQHANLFINLFSMMLGSGMPELQSFDDIAYIRKTLALDKTEQEALEYFTKQMNDAHHGGWTTKMDWIFHTIKQHALN DIP protein P42337 original sequence version: 123 D593283B416ABFD0 pgfrb_mouse Platelet-derived growth factor receptor beta CD140 antigen-like family member B Pdgfrb Pdgfr Beta-type platelet-derived growth factor receptor Pdgfr1 Platelet-derived growth factor receptor 1 Beta platelet-derived growth factor receptor protein protein mouse Mus musculus Mouse MGLPGVIPALVLRGQLLLSVLWLLGPQTSRGLVITPPGPEFVLNISSTFVLTCSGSAPVMWEQMSQVPWQEAAMNQDGTFSSVLTLTNVTGGDTGEYFCVYNNSLGPELSERKRIYIFVPDPTMGFLPMDSEDLFIFVTDVTETTIPCRVTDPQLEVTLHEKKVDIPLHVPYDHQRGFTGTFEDKTYICKTTIGDREVDSDTYYVYSLQVSSINVSVNAVQTVVRQGESITIRCIVMGNDVVNFQWTYPRMKSGRLVEPVTDYLFGVPSRIGSILHIPTAELSDSGTYTCNVSVSVNDHGDEKAINISVIENGYVRLLETLGDVEIAELHRSRTLRVVFEAYPMPSVLWLKDNRTLGDSGAGELVLSTRNMSETRYVSELILVRVKVSEAGYYTMRAFHEDDEVQLSFKLQVNVPVRVLELSESHPANGEQTIRCRGRGMPQPNVTWSTCRDLKRCPRKLSPTPLGNSSKEESQLETNVTFWEEDQEYEVVSTLRLRHVDQPLSVRCMLQNSMGGDSQEVTVVPHSLPFKVVVISAILALVVLTVISLIILIMLWQKKPRYEIRWKVIESVSSDGHEYIYVDPVQLPYDSTWELPRDQLVLGRTLGSGAFGQVVEATAHGLSHSQATMKVAVKMLKSTARSSEKQALMSELKIMSHLGPHLNVVNLLGACTKGGPIYIITEYCRYGDLVDYLHRNKHTFLQRHSNKHCPPSAELYSNALPVGFSLPSHLNLTGESDGGYMDMSKDESIDYVPMLDMKGDIKYADIESPSYMAPYDNYVPSAPERTYRATLINDSPVLSYTDLVGFSYQVANGMDFLASKNCVHRDLAARNVLICEGKLVKICDFGLARDIMRDSNYISKGSTFLPLKWMAPESIFNSLYTTLSDVWSFGILLWEIFTLGGTPYPELPMNDQFYNAIKRGYRMAQPAHASDEIYEIMQKCWEEKFETRPPFSQLVLLLERLLGEGYKKKYQQVDEEFLRSDHPAILRSQARFPGIHSLRSPLDTSSVLYTAVQPNESDNDYIIPLPDPKPDVADEGLPEGSPSLASSTLNEVNTSSTISCDSPLELQEEPQQAEPEAQLEQPQDSGCPGPLAEAEDSFL DIP protein P05622 original sequence version: 166 DIP protein P05622 original sequence version: 148 D08FC7C42B77B794 gtp guanosine triphosphate - GTP 5'-GTP GTP Guanosine 5'-triphosphate guanosine 5'-triphosphoric acid guanosine triphosphate GUANOSINE-5'-TRIPHOSPHATE H4gtp guanosine 5'-(tetrahydrogen triphosphate) small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) InChI=1/C10H16N5O14P3/c11-10-13-7-4(8(18)14-10)12-2-15(7)9-6(17)5(16)3(27-9)1-26-31(22,23)29-32(24,25)28-30(19,20)21/h2-3,5-6,9,16-17H,1H2,(H,22,23)(H,24,25)(H2,19,20,21)(H3,11,13,14,18)/t3-,5-,6-,9-/m1/s1/f/h14,19-20,22,24H,11H2 InChI=1/C10H16N5O14P3/c11-10-13-7-4(8(18)14-10)12-2-15(7)9-6(17)5(16)3(27-9)1-26-31(22,23)29-32(24,25)28-30(19,20)21/h2-3,5-6,9,16-17H,1H2,(H,22,23)(H,24,25)(H2,19,20,21)(H3,11,13,14,18)/t3-,5-,6-,9-/m1/s1/f/h14,19-20,22,24H,11H2 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 p85a_bovin Phosphatidylinositol 3-kinase regulatory subunit alpha Phosphatidylinositol 3-kinase 85 kDa regulatory subunit alpha PIK3R1 protein protein bovin Bos taurus (Bovine) Bovine MSAEGYQYRALYDYKKEREEDIDLHLGDILTVNKGSLVALGFSDGQEAKPEEIGWLNGYNETTGERGDFPGTYVEYIGRKKISPPTPKPRPPRPLPVAPGPSKTEADSEQQASTLPDLAEQFAPPDVAPPLLIKLVEAIEKKGLECSTLYRTQSSSNPAELRQLLDCDTASLDLEMFDVHVLADAFKRYLLDLPNPVIPVAVSSELISLAPEVQSSEEYIQLLKKLIRSPSIPHQYWLTLQYLLKHFFKLSQTSSKNLLNARVLSELFSPLLFRFPAASSENTEHLIKIIEILISTEWNERQPAPALPPKPPKPTTVANNGMNNNMSLQDAEWYWGDISREEVNEKLRDTADGTFLVRDASTKMHGDYTLTLRKGGNNKLIKIFHRDGKYGFSDPLTFNSVVELINHYRNESLAQYNPKLDVKLLYPVSKYQQDQVVKEDNIEAVGKKLHEYNTQFQEKSREYDRLYEDYTRTSQEIQMKRTAIEAFNETIKIFEEQCQTQERYSKEYIEKFKREGNETEIQRIMHNYEKLKSRISEIVDSRRRLEEDLKKQAAEYREIDKRMNSIKPDLIQLRKTRDQYLMWLTQKGVRQKKLNEWLGNENTEDQYSLVEDDEDLPHHDEKTWNVGSSNRNKAENLLRGKRDGTFLVRESSKQGCYACSVVVDGEVKHCVINKTATGYGFAEPYNLYSSLKELVLHYQHTSLVQHNDSLNVTLAYPVYAQQRR DIP protein P23727 original sequence version: 108 EBDF6E754BBF7321 rab4a_human Ras-related protein Rab-4A RAB4A RAB4 protein protein human Homo sapiens Human MSQTAMSETYDFLFKFLVIGNAGTGKSCLLHQFIEKKFKDDSNHTIGVEFGSKIINVGGKYVKLQIWDTAGQERFRSVTRSYYRGAAGALLVYDITSRETYNALTNWLTDARMLASQNIVIILCGNKKDLDADREVTFLEASRFAQENELMFLETSALTGENVEEAFVQCARKILNKIESGELDPERMGSGIQYGDAALRQLRSPRRAQAPNAQECGC DIP protein P20338 original sequence version: 151 983D65E1162741B0 rab6a_human Ras-related protein Rab-6A RAB6A RAB6 protein protein human Homo sapiens Human MSTGGDFGNPLRKFKLVFLGEQSVGKTSLITRFMYDSFDNTYQATIGIDFLSKTMYLEDRTVRLQLWDTAGQERFRSLIPSYIRDSTVAVVVYDITNVNSFQQTTKWIDDVRTERGSDVIIMLVGNKTDLADKRQVSIEEGERKAKELNVMFIETSAKAGYNVKQLFRRVAAALPGMESTQDRSREDMIDIKLEKPQEQPVSEGGCSC C9A3C2DDBF6C3E9E rac1_human Ras-related C3 botulinum toxin substrate 1 RAC1 MIG5 p21-Rac1 Ras-like protein TC25 Cell migration-inducing gene 5 protein TC25 protein protein human Homo sapiens Human MQAIKCVVVGDGAVGKTCLLISYTTNAFPGEYIPTVFDNYSANVMVDGKPVNLGLWDTAGQEDYDRLRPLSYPQTDVFLICFSLVSPASFENVRAKWYPEVRHHCPNTPIILVGTKLDLRDDKDTIEKLKEKKLTPITYPQGLAMAKEIGAVKYLECSALTQRGLKTVFDEAIRAVLCPPPVKKRKRKCLLL ACEDF83A45E5EA67 cdc42_human Cell division control protein 42 homolog CDC42 G25K GTP-binding protein protein protein human Homo sapiens Human MQTIKCVVVGDGAVGKTCLLISYTTNKFPSEYVPTVFDNYAVTVMIGGEPYTLGLFDTAGQEDYDRLRPLSYPQTDVFLVCFSVVSPSSFENVKEKWVPEITHHCPKTPFLLVGTQIDLRDDPSTIEKLAKNKQKPITPETAEKLARDLKAVKYVECSALTQKGLKNVFDEAILAALEPPEPKKSRRCVLL 51A437E22A4D8FFF rab5a-gtp 5 7 enzyme enzyme unspecified role unspecified role ecobb Escherichia coli (strain B / BL21) 10 enzyme target enzyme target substrate substrate unspecified role unspecified role 12 stimulator stimulator unspecified role unspecified role P23727:p.Arg274Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain R A Wed May 02 20:11:52 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg274ala' Tue Jun 30 19:21:29 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg274Ala' P23727:p.Arg151Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain R A Wed May 02 19:23:15 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg151ala' Tue Jun 30 17:29:03 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg151Ala' region BH domain required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecobb Escherichia coli (strain B / BL21) enzymatic reaction enzymatic reaction Biochemical reaction Biochemical reaction 1, 2 The addition of p85alpha to the assay substantially accelerated Rab5-mediated hydrolysis of the bound [alpha-32P]GTP to [alpha-32P]GDP, suggesting that p85 stimulates GTP hydrolysis by Rab5 and is not simply promoting nucleotide exchange. The addition of p85 to the assay did substantially accelerate the Rab5-mediated hydrolysis of the bound [-32P]GTP to [-32P]GDP, p85alpha - Addition of p85 to the assay substantially accelerated Rab5-mediated hydrolysis of the bound [alpha-32P]GTP to [alpha-32P]GDP. Stimulation increased as more p85alpha was added. The BH domain of p85alpha contained substantial Rab5 GAP activity, as deletion mutants lacking this domain showed little or no GAP activity towards Rab5. rab5a-pik3r1 3 7 unspecified role unspecified role bait bait ecobb Escherichia coli (strain B / BL21) 12 unspecified role unspecified role prey prey ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction 3 A The p85 protein bound directly to Rab5 in a concentration-dependent manner This interaction was similar in the absence of nucleotide, or in the presence of GDP or GTPgammaS (a non-hydrolyzable form of GTP). rab5a-pik3r1-1 6 12 unspecified role unspecified role prey prey ecobb Escherichia coli (strain B / BL21) 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 ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction 3 B, C Purified cleaved p85 pro-tein bound best to Rab5 (no nucleotide), followed by Rab5-GTPS. Significant amounts of p85 also bound to the transition state analogue conformation of Rab5 (GDP-AlF4), and to Rab5-GDP. We found similar results using a pull-down assay with two Rab5 mutants. The Rab5S34N mutant preferentially binds GDP, whereas the Rab5Q79L mutant is unable to hydrolyze bound GTP. Purified cleaved p85 bound to both GSTRab5S34N and GST-RabQ79L preloaded with GDP and GTP, respectively, which were immobilized on glutathione-Sepharose beads Purified cleaved p85 protein bound best to Rab5 (no nucleotide), followed by Rab5-GTPgammaS. Significant amounts of p85 also bound to the transition state analogue conformation of Rab5 (GDP-AlF4), and to Rab5-GDP.The Rab5S34N mutant preferentially binds GDP, whereas the Rab5Q79L mutant is unable to hydrolyze bound GTP. Purified cleaved p85 bound to both GSTRab5S34N and GST-RabQ79L preloaded with GDP and GTP. rab5a-pik3r1-2 6 7 unspecified role unspecified role bait bait P20339:p.Gln79Leu mutation mutation mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Q L Wed May 02 15:24:59 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gln79leu' Tue Jun 30 19:41:44 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gln79Leu' P20339:p.Ser34Asn mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain S N Wed May 02 18:55:30 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser34asn' Tue Jun 30 18:27:37 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser34Asn' 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 ecobb Escherichia coli (strain B / BL21) 12 unspecified role unspecified role prey prey region p85alphaDeltaBH 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 ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction 3 D The p85BH mutant only bound Rab5Q79L-GTP Rab5Q79L was loaded with GTP, whereas Rab5S34N was loaded with GDP. rab5a-pik3r1-3 6 12 unspecified role unspecified role prey prey P23727:p.Arg151Ala mutation mutation mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain R A Wed May 02 17:10:59 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg151ala' Tue Jun 30 20:12:36 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg151Ala' ecobb Escherichia coli (strain B / BL21) 7 unspecified role unspecified role bait bait P20339:p.Gln79Leu mutation mutation mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Q L Wed May 02 20:16:23 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gln79leu' Tue Jun 30 18:08:07 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gln79Leu' P20339:p.Ser34Asn mutation mutation mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain S N Wed May 02 19:08:28 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser34asn' Tue Jun 30 18:15:07 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser34Asn' 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 ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction 3 D Rab5Q79L was loaded with GTP, whereas Rab5S34N was loaded with GDP. rab5a-pik3r1-4 6 12 unspecified role unspecified role prey prey P23727:p.Arg274Ala mutation mutation mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain R A Wed May 02 15:11:23 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'arg274ala' Tue Jun 30 18:28:05 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Arg274Ala' ecobb Escherichia coli (strain B / BL21) 7 unspecified role unspecified role bait bait P20339:p.Ser34Asn mutation mutation mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain S N Wed May 02 17:54:23 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser34asn' Tue Jun 30 16:50:23 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser34Asn' P20339:p.Gln79Leu mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Q L Wed May 02 16:19:27 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gln79leu' Tue Jun 30 18:35:47 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gln79Leu' 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 ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction 3 D the p85R274A mutant only bound Rab5S34N-GDP. Rab5Q79L was loaded with GTP, whereas Rab5S34N was loaded with GDP. rab4a-gtp 5 13 enzyme enzyme unspecified role unspecified role ecobb Escherichia coli (strain B / BL21) 10 enzyme target enzyme target substrate substrate unspecified role unspecified role 12 stimulator stimulator unspecified role unspecified role ecobb Escherichia coli (strain B / BL21) enzymatic reaction enzymatic reaction Biochemical reaction Biochemical reaction 4 A The p85 protein displayed similar GAP activity toward Rab5 and Rab4, and of the Rab proteins tested, these were the GTPases most stimulated by p85 rab6a-gtp-1 5 10 enzyme target enzyme target substrate substrate unspecified role unspecified role 14 enzyme enzyme unspecified role unspecified role ecobb Escherichia coli (strain B / BL21) 12 stimulator stimulator unspecified role unspecified role ecobb Escherichia coli (strain B / BL21) enzymatic reaction enzymatic reaction Biochemical reaction Biochemical reaction 4 A The GTPase activity of Rab6 was also stimulated by p85 The GTPase activity of Rab6 was stimulated by p85alpha to a lesser degree than Rab5 or Rab4. rac1-gtp 5 15 stimulator stimulator unspecified role unspecified role ecobb Escherichia coli (strain B / BL21) 12 stimulator stimulator unspecified role unspecified role ecobb Escherichia coli (strain B / BL21) 10 enzyme target enzyme target substrate substrate unspecified role unspecified role enzymatic reaction enzymatic reaction Biochemical reaction Biochemical reaction 4 B We observed that p85 did have GAP activity toward both Rac1 and Cdc42 p85 had a similar level of GAP activity toward Rac1 as Rab4 and Rab5. cdc42-gtp 5 12 stimulator stimulator unspecified role unspecified role ecobb Escherichia coli (strain B / BL21) 10 enzyme target enzyme target substrate substrate unspecified role unspecified role 16 enzyme enzyme unspecified role unspecified role ecobb Escherichia coli (strain B / BL21) enzymatic reaction enzymatic reaction Biochemical reaction Biochemical reaction 4 B We observed that p85 did have GAP activity toward both Rac1 and Cdc42 p85 had a similar level of GAP activity toward Cdc42 as Rab4 and Rab5. pik3r1-pik3ca-3 1 8 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 12 unspecified role unspecified role bait bait region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association 5 These results indicated that all of the FLAGp85 proteins (wild type, BH, R274A, and R151A mutants) can associate with the Myc-p110 protein and its encoded PI3K activity. All of the FLAG-p85alpha proteins (wild type, DeltaBH, R274A, and R151A mutants) can associate with the Myc-p110alpha. P42337; P23727 transfection pdgfrb-pik3r1 2 11 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait physical association physical association 6 A The PDGFR typically binds p85 in a PDGF-dependent manner, as shown by the presence of p85 in anti-PDGFR immunoprecipitates from PDGF-treated NIH 3T3 cell lysates PDGF BB - PDGF BB treatment stimulated the interaction before it decreased again with time of treatment. pdgfrb-pik3r1-1 4 12 unspecified role unspecified role prey prey region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 9 unspecified role unspecified role bait bait physical association physical association 6 A we find that anti-PDGFR immunoprecipitates contain FLAG-p85 proteins (both wild type and BH mutants BH and R274A) after PDGF stimulation. PDGFR coimmunoprecipitates wild-type FLAG-p85 and also DeltaBH and R274A. P23727 transfection PDGF BB - PDGF BB treatment stimulates this interaction. The interaction decreases with time of treatment for FLAG-p85DeltaBH and more markedly for the wild-type protein, but not for FLAG-p85R274A.