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/ EphA2 mediates ligand-dependent inhibition and ligand-independent promotion of cell migration and invasion via a reciprocal regulatory loop with Akt. EphA2 mediates ligand-dependent inhibition and ligand-independent promotion of cell migration and invasion via a reciprocal regulatory loop with Akt. Cancer Cell (1535-6108) 2009 imex curation Miao H., Li DQ., Mukherjee A., Guo H., Petty A., Cutter J., Basilion JP., Sedor J., Wu J., Danielpour D., Sloan AE., Cohen ML., Wang B. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer U373 cells: this was inferred from the text: not stated on the image/image legend. We do not have "Expressed in" species for ephrin-A1-Fc. FBS Additional detail of constructs supplied by the author. hxm35@case.edu 29-Mar-2010: Contacted by JYOTI. orchard Only protein-protein interactions 5 human-u373_mg Human glioblastoma astrocytoma u373_mg Human glioblastoma astrocytoma anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Miao H., Li DQ., Mukherjee A., Guo H., Petty A., Cutter J., Basilion JP., Sedor J., Wu J., Danielpour D., Sloan AE., Cohen ML., Wang B. Apoptosis - Interactions involving proteins with a function related to apoptosis U373 cells: this was inferred from the text: not stated on the image/image legend. We do not have "Expressed in" species for ephrin-A1-Fc. FBS Additional detail of constructs supplied by the author. hxm35@case.edu hxm35@case.edu Accepted 2010-MAR-26 by ORCHARD. 29-Mar-2010: Contacted by JYOTI. Cancer Cell (1535-6108) 2009 orchard Only protein-protein interactions 5 imex curation EphA2 mediates ligand-dependent inhibition and ligand-independent promotion of cell migration and invasion via a reciprocal regulatory loop with Akt. EphA2 mediates ligand-dependent inhibition and ligand-independent promotion of cell migration and invasion via a reciprocal regulatory loop with Akt. Cancer Cell (1535-6108) 2009 imex curation Miao H., Li DQ., Mukherjee A., Guo H., Petty A., Cutter J., Basilion JP., Sedor J., Wu J., Danielpour D., Sloan AE., Cohen ML., Wang B. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer U373 cells: this was inferred from the text: not stated on the image/image legend. We do not have "Expressed in" species for ephrin-A1-Fc. FBS Additional detail of constructs supplied by the author. hxm35@case.edu 29-Mar-2010: Contacted by JYOTI. orchard Only protein-protein interactions 5 human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Miao H., Li DQ., Mukherjee A., Guo H., Petty A., Cutter J., Basilion JP., Sedor J., Wu J., Danielpour D., Sloan AE., Cohen ML., Wang B. Apoptosis - Interactions involving proteins with a function related to apoptosis U373 cells: this was inferred from the text: not stated on the image/image legend. We do not have "Expressed in" species for ephrin-A1-Fc. FBS Additional detail of constructs supplied by the author. hxm35@case.edu hxm35@case.edu Accepted 2010-MAR-26 by ORCHARD. 29-Mar-2010: Contacted by JYOTI. Cancer Cell (1535-6108) 2009 orchard Only protein-protein interactions 5 imex curation EphA2 mediates ligand-dependent inhibition and ligand-independent promotion of cell migration and invasion via a reciprocal regulatory loop with Akt. EphA2 mediates ligand-dependent inhibition and ligand-independent promotion of cell migration and invasion via a reciprocal regulatory loop with Akt. Cancer Cell (1535-6108) 2009 imex curation Miao H., Li DQ., Mukherjee A., Guo H., Petty A., Cutter J., Basilion JP., Sedor J., Wu J., Danielpour D., Sloan AE., Cohen ML., Wang B. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer U373 cells: this was inferred from the text: not stated on the image/image legend. We do not have "Expressed in" species for ephrin-A1-Fc. FBS Additional detail of constructs supplied by the author. hxm35@case.edu 29-Mar-2010: Contacted by JYOTI. orchard Only protein-protein interactions 5 human-pc3m Human PC3M Caucasian prostate adenocarcinoma pc3m Human PC3M Caucasian prostate adenocarcinoma PC-3M was isolated from a liver metastasis produced by the intrasplenic injection of human PC-3 prostate cancer cells into nude mice. PC-3M cells have "Hyperactive Akt" and are PTEN null. anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Miao H., Li DQ., Mukherjee A., Guo H., Petty A., Cutter J., Basilion JP., Sedor J., Wu J., Danielpour D., Sloan AE., Cohen ML., Wang B. Apoptosis - Interactions involving proteins with a function related to apoptosis U373 cells: this was inferred from the text: not stated on the image/image legend. We do not have "Expressed in" species for ephrin-A1-Fc. FBS Additional detail of constructs supplied by the author. hxm35@case.edu hxm35@case.edu Accepted 2010-MAR-26 by ORCHARD. 29-Mar-2010: Contacted by JYOTI. Cancer Cell (1535-6108) 2009 orchard Only protein-protein interactions 5 imex curation epha2_human Ephrin type-A receptor 2 Tyrosine-protein kinase receptor ECK Epithelial cell kinase EPHA2 ECK protein protein human Homo sapiens Human MELQAARACFALLWGCALAAAAAAQGKEVVLLDFAAAGGELGWLTHPYGKGWDLMQNIMNDMPIYMYSVCNVMSGDQDNWLRTNWVYRGEAERIFIELKFTVRDCNSFPGGASSCKETFNLYYAESDLDYGTNFQKRLFTKIDTIAPDEITVSSDFEARHVKLNVEERSVGPLTRKGFYLAFQDIGACVALLSVRVYYKKCPELLQGLAHFPETIAGSDAPSLATVAGTCVDHAVVPPGGEEPRMHCAVDGEWLVPIGQCLCQAGYEKVEDACQACSPGFFKFEASESPCLECPEHTLPSPEGATSCECEEGFFRAPQDPASMPCTRPPSAPHYLTAVGMGAKVELRWTPPQDSGGREDIVYSVTCEQCWPESGECGPCEASVRYSEPPHGLTRTSVTVSDLEPHMNYTFTVEARNGVSGLVTSRSFRTASVSINQTEPPKVRLEGRSTTSLSVSWSIPPPQQSRVWKYEVTYRKKGDSNSYNVRRTEGFSVTLDDLAPDTTYLVQVQALTQEGQGAGSKVHEFQTLSPEGSGNLAVIGGVAVGVVLLLVLAGVGFFIHRRRKNQRARQSPEDVYFSKSEQLKPLKTYVDPHTYEDPNQAVLKFTTEIHPSCVTRQKVIGAGEFGEVYKGMLKTSSGKKEVPVAIKTLKAGYTEKQRVDFLGEAGIMGQFSHHNIIRLEGVISKYKPMMIITEYMENGALDKFLREKDGEFSVLQLVGMLRGIAAGMKYLANMNYVHRDLAARNILVNSNLVCKVSDFGLSRVLEDDPEATYTTSGGKIPIRWTAPEAISYRKFTSASDVWSFGIVMWEVMTYGERPYWELSNHEVMKAINDGFRLPTPMDCPSAIYQLMMQCWQQERARRPKFADIVSILDKLIRAPDSLKTLADFDPRVSIRLPSTSGSEGVPFRTVSEWLESIKMQQYTEHFMAAGYTAIEKVVQMTNDDIKRIGVRLPGHQKRIAYSLLGLKDQVNTVGIPI DIP protein P29317 original sequence version: 111 DIP protein P29317 original sequence version: 126 DIP protein P29317 original sequence version: 141 DIP protein P29317 original sequence version: 146 845D7E1BBCCAACCC efna1_human Ephrin-A1 EPH-related receptor tyrosine kinase ligand 1 Immediate early response protein B61 Tumor necrosis factor alpha-induced protein 4 EFNA1 EPLG1 LERK1 TNFAIP4 protein protein human Homo sapiens Human MEFLWAPLLGLCCSLAAADRHTVFWNSSNPKFRNEDYTIHVQLNDYVDIICPHYEDHSVADAAMEQYILYLVEHEEYQLCQPQSKDQVRWQCNRPSAKHGPEKLSEKFQRFTPFTLGKEFKEGHSYYYISKPIHQHEDRCLRLKVTVSGKITHSPQAHDNPQEKRLAADDPEVRVLHSIGHSAAPRLFPLAWTVLLLPLLLLQTP DIP protein P20827 original sequence version: 161 DIP protein P20827 original sequence version: 96 DIP protein P20827 original sequence version: 111 DIP protein P20827 original sequence version: 116 EFF026BF4C12461F efa1-epha2-2 1 4 unspecified role unspecified role prey prey 5 unspecified role unspecified role bait bait ephrin-a1-fc Ephrin-A1-Fc fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Ephrin-A1-Fc is a recombinant protein composed of ephrin-A1 and Fc (human IgG1 heavy chain) moieties. It binds to all EphA kinases. An antibody recognizing the consensus Akt substrate sites (Akt-pSub). detected a strong band (at 125 kDa) indicating that EphA2 could be a substrate for Akt. Ephrin-A1 cotreatment led to complete inhibition of EphA2 phosphorylation at Akt substrate sites, which correlated with the abolishment of Akt activation. Serum-induced EphA2 phosphorylation at Akt substrate sites is abolished by pretreating cells with LY294002. Fig 1H, Fig 1I. PI3K inhibitor LY294002 efa1-epha2-1 1 4 unspecified role unspecified role prey prey 5 unspecified role unspecified role bait bait ephrin-a1-fc Ephrin-A1-Fc fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Ephrin-A1-Fc is a recombinant protein composed of ephrin-A1 and Fc (human IgG1 heavy chain) moieties. It binds to all EphA kinases. Fig 1J. Serum-induced S/T phosphorylation of EphA2 was attenuated by the expression of DN Akt. efna1-epha2-3 2 5 unspecified role unspecified role bait bait ephrin-a1-fc Ephrin-A1-Fc fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 4 unspecified role unspecified role prey prey P29317:p.Ser897Asp 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 19:53:58 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser897asp' Tue Jun 30 19:46:17 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser897Asp' P29317:p.Ser897Ala mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain S A Wed May 02 15:46:19 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser897ala' Tue Jun 30 16:40:01 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser897Ala' physical association physical association Ephrin-A1-Fc is a recombinant protein composed of ephrin-A1 and Fc (human IgG1 heavy chain) moieties. It binds to all EphA kinases. Both S897A and S897D mutations completely abolish the serum-induced phosphorylation of EphA2 on Akt substrate sites. Fig 2B, 2D. efna1-epha2-1 2 5 unspecified role unspecified role bait bait ephrin-a1-fc Ephrin-A1-Fc fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 4 unspecified role unspecified role prey prey ser-897 (possible) opser O-phospho-L-serine (2S)-2-amino-3-(phosphonooxy)propanoic acid 2-amino-3-hydroxypropanoic acid 3-phosphate 2-azanyl-3-(phosphonooxy)propanoic acid ACT_SITE Phosphoserine intermediate MOD_RES Phosphoserine O-phospho-L-serine O-phosphonoserine O-phosphorylated L-serine O3-phosphoserine serine phosphate ester OPSer Phospho Phospho Seryl Phosphorylation phosphorylations certain certain sequence position certain certain certain sequence position certain physical association physical association Ephrin-A1-Fc is a recombinant protein composed of ephrin-A1 and Fc (human IgG1 heavy chain) moieties. It binds to all EphA kinases. A robust pS897 band comigrating with EphA2 was also detected in EphA2 immunoprecipitates, but not in cells expressing S897A-EphA2. Fig 5B. efa1-epha2 3 4 unspecified role unspecified role prey prey 5 unspecified role unspecified role bait bait ephrin-a1-fc Ephrin-A1-Fc fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Ephrin-A1-Fc is a recombinant protein composed of ephrin-A1 and Fc (human IgG1 heavy chain) moieties. It binds to all EphA kinases. Fig S4. Hyperactive Akt was also observed in the PTEN null PC-3M cells. EphA2 kinase activation by ephrin-A1 stimulation inhibits Akt activation in migrating PC-3M cells.