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 RAP1 guanine nucleotide exchange factor Epac2 couples cyclic AMP and Ras signals at the plasma membrane. The RAP1 guanine nucleotide exchange factor Epac2 couples cyclic AMP and Ras signals at the plasma membrane. J. Biol. Chem. (0021-9258) 2006 imex curation Li Y., Asuri S., Rebhun JF., Castro AF., Paranavitana NC., Quilliam LA. BioCreative - Critical Assessment of Information Extraction systems in Biology Ref. 13 states that H-ras, N-ras and K-ras are human ras genes. A PubMed search against Gaudriault (one of the donors of the pcDNA3-Epac2) and Epac reveals that this construct contains the human sequence of Epac2. Note the co-ip for the mutants of N-Ras and K-Ras were not included as the wild-type proteins were not used anywhere. lquillia@iupui.edu 24-JAN-2007: Contacted by JYOTI. 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 western blot western blot Immuno blot Immuno blot Li Y., Asuri S., Rebhun JF., Castro AF., Paranavitana NC., Quilliam LA. BioCreative - Critical Assessment of Information Extraction systems in Biology Ref. 13 states that H-ras, N-ras and K-ras are human ras genes. A PubMed search against Gaudriault (one of the donors of the pcDNA3-Epac2) and Epac reveals that this construct contains the human sequence of Epac2. Note the co-ip for the mutants of N-Ras and K-Ras were not included as the wild-type proteins were not used anywhere. lquillia@iupui.edu Accepted 2006-SEP-1 by DAVET. J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation The RAP1 guanine nucleotide exchange factor Epac2 couples cyclic AMP and Ras signals at the plasma membrane. The RAP1 guanine nucleotide exchange factor Epac2 couples cyclic AMP and Ras signals at the plasma membrane. J. Biol. Chem. (0021-9258) 2006 imex curation Li Y., Asuri S., Rebhun JF., Castro AF., Paranavitana NC., Quilliam LA. BioCreative - Critical Assessment of Information Extraction systems in Biology Ref. 13 states that H-ras, N-ras and K-ras are human ras genes. A PubMed search against Gaudriault (one of the donors of the pcDNA3-Epac2) and Epac reveals that this construct contains the human sequence of Epac2. Note the co-ip for the mutants of N-Ras and K-Ras were not included as the wild-type proteins were not used anywhere. lquillia@iupui.edu 24-JAN-2007: Contacted by JYOTI. Only protein-protein interactions in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Li Y., Asuri S., Rebhun JF., Castro AF., Paranavitana NC., Quilliam LA. BioCreative - Critical Assessment of Information Extraction systems in Biology Ref. 13 states that H-ras, N-ras and K-ras are human ras genes. A PubMed search against Gaudriault (one of the donors of the pcDNA3-Epac2) and Epac reveals that this construct contains the human sequence of Epac2. Note the co-ip for the mutants of N-Ras and K-Ras were not included as the wild-type proteins were not used anywhere. lquillia@iupui.edu Accepted 2006-SEP-1 by DAVET. 24-JAN-2007: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation The RAP1 guanine nucleotide exchange factor Epac2 couples cyclic AMP and Ras signals at the plasma membrane. The RAP1 guanine nucleotide exchange factor Epac2 couples cyclic AMP and Ras signals at the plasma membrane. J. Biol. Chem. (0021-9258) 2006 imex curation Li Y., Asuri S., Rebhun JF., Castro AF., Paranavitana NC., Quilliam LA. BioCreative - Critical Assessment of Information Extraction systems in Biology Ref. 13 states that H-ras, N-ras and K-ras are human ras genes. A PubMed search against Gaudriault (one of the donors of the pcDNA3-Epac2) and Epac reveals that this construct contains the human sequence of Epac2. Note the co-ip for the mutants of N-Ras and K-Ras were not included as the wild-type proteins were not used anywhere. lquillia@iupui.edu 24-JAN-2007: Contacted by JYOTI. Only protein-protein interactions human-h1299 Homo sapiens lung lymph node carcinoma h1299 Human lung lymph node carcinoma p53 null. These cells have also been reported to secrete the peptide hormone neuromedin B (NMB), but not gastrin releasing peptide (GRP). Also known as NCI H1299 or CRL-5803. http://www.lgcpromochem.com/atcc/ ATCC number: CRL-5803. ATCC number of the cell line is CRL-5803. confocal microscopy confocal microscopy predetermined predetermined participant predetermined Li Y., Asuri S., Rebhun JF., Castro AF., Paranavitana NC., Quilliam LA. BioCreative - Critical Assessment of Information Extraction systems in Biology Ref. 13 states that H-ras, N-ras and K-ras are human ras genes. A PubMed search against Gaudriault (one of the donors of the pcDNA3-Epac2) and Epac reveals that this construct contains the human sequence of Epac2. Note the co-ip for the mutants of N-Ras and K-Ras were not included as the wild-type proteins were not used anywhere. lquillia@iupui.edu Accepted 2006-SEP-1 by DAVET. 24-JAN-2007: Contacted by JYOTI. J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation rash_human GTPase HRas HRAS HRAS1 Transforming protein p21 p21ras H-Ras-1 c-H-ras Ha-Ras protein protein human Homo sapiens Human MTEYKLVVVGAGGVGKSALTIQLIQNHFVDEYDPTIEDSYRKQVVIDGETCLLDILDTAGQEEYSAMRDQYMRTGEGFLCVFAINNTKSFEDIHQYREQIKRVKDSDDVPMVLVGNKCDLAARTVESRQAQDLARSYGIPYIETSAKTRQGVEDAFYTLVREIRQHKLRKLNPPDESGPGCMSCKCVLS DIP protein P01112 original sequence version: 91 DIP protein P01112 original sequence version: 106 DIP protein P01112 original sequence version: 144 DIP protein P01112 original sequence version: 118 DIP protein P01112 original sequence version: 143 DIP protein P01112 original sequence version: 152 DIP protein P01112-1 original sequence version: 159 DIP protein P01112 original sequence version: 170 DIP protein P01112-1 original sequence version: 185 DIP protein P01112 original sequence version: 192 DIP protein P01112-1 original sequence version: 196 DIP protein P01112-1 original sequence version: 202 EE6DC2D933E2856A rpgf4_mouse Rap guanine nucleotide exchange factor 4 cAMP-regulated guanine nucleotide exchange factor II Exchange factor directly activated by cAMP 2 Exchange protein directly activated by cAMP 2 cAMP-dependent Rap1 guanine-nucleotide exchange factor Rapgef4 Cgef2 Epac2 protein protein mouse Mus musculus Mouse MVAAHAAHSQSSAEWIACLDKRPLERSSEDVDIIFTRLKGVKAFEKFHPNLLRQICLCGYYENLEKGITLFRQGDIGTNWYAVLAGSLDVKVSETSSHQDAVTICTLGIGTAFGESILDNTPRHATIVTRESSELLRIEQEDFKALWEKYRQYMAGLLAPPYGVMETGSNNDRIPDKENTPLIEPHVPLRPAHTITKVPSEKILRAGKILRIAILSRAPHMIRDRKYHLKTYRQCCVGTELVDWMIQQTSCVHSRTQAVGMWQVLLEDGVLNHVDQERHFQDKYLFYRFLDDEREDAPLPTEEEKKECDEELQDTMLLLSQMGPDAHMRMILRKPPGQRTVDDLEIIYDELLHIKALSHLSTTVKRELAGVLIFESHAKGGTVLFNQGEEGTSWYIILKGSVNVVIYGKGVVCTLHEGDDFGKLALVNDAPRAASIVLREDNCHFLRVDKEDFNRILRDVEANTVRLKEHDQDVLVLEKVPAGNRAANQGNSQPQQKYTVMSGTPEKILEHFLETIRLEPSLNEATDSVLNDFVMMHCVFMPNTQLCPALVAHYHAQPSQGTEQERMDYALNNKRRVIRLVLQWAAMYGDLLQEDDVAMAFLEEFYVSVSDDARMMAAFKEQLPELEKIVKQISEDAKAPQKKHKVLLQQFNTGDERAQKRQPIRGSDEVLFKVYCIDHTYTTIRVPVAASVKEVISAVADKLGSGEGLIIVKMNSGGEKVVLKSNDVSVFTTLTINGRLFACPREQFDSLTPLPEQEGPTTGTVGTFELMSSKDLAYQMTTYDWELFNCVHELELIYHTFGRHNFKKTTANLDLFLRRFNEIQFWVVTEVCLCSQLSKRVQLLKKFIKIAAHCKEYKNLNSFFAIVMGLSNVAVSRLALTWEKLPSKFKKFYAEFESLMDPSRNHRAYRLTAAKLEPPLIPFMPLLIKDMTFTHEGNKTFIDNLVNFEKMRMIANTARTVRYYRSQPFNPDAAQANKNHQDVRSYVRQLNVIDNQRTLSQMSHRLEPRRP 449BC537475B03AA hras-rapgef4 2 4 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) 5 unspecified role unspecified role prey prey n-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range region Epac2deltaRA required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain chlae-cos_1 Chlorocebus aethiops COS-1 cells Grivet cos_1 CV-1 Simian cells (cercopithecus aethiops) transformed by SV40 physical association physical association Epac2 specifically bound GTP S-loaded H-Ras. 2 A, B GTPgammaS-loaded GST-H-Ras participated in the interaction but not GDP-loaded protein. Note that the authors refer to COS cells. It is assumed that these are Cos-1 cells. hras-rapgef4-1 1 5 unspecified role unspecified role prey prey region Epac2deltaRA required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain n-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range 4 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 P01112:p.Gln61Leu mutation mutation mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Q L Wed May 02 20:09:50 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gln61leu' Tue Jun 30 18:01:11 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gln61Leu' P01112:p.Gly12Val mutation mutation mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain G V Wed May 02 17:56:30 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly12val' Tue Jun 30 15:54:14 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gly12Val' P01112:p.[Gly12Val;Thr35Ser] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain G V certain certain sequence position certain certain certain sequence position certain T S Wed May 02 19:52:36 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly12val,thr35ser' Tue Jun 30 18:23:09 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Gly12Val;Thr35Ser]' P01112:p.[Gly12Val;Glu37Gly] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain G V certain certain sequence position certain certain certain sequence position certain E G Wed May 02 14:38:16 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly12val,glu37gly' Tue Jun 30 17:10:07 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Gly12Val;Glu37Gly]' P01112:p.[Gly12Val;Tyr40Cys] mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Y C certain certain sequence position certain certain certain sequence position certain G V Fri Jul 22 11:07:34 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'tyr40cys,gly12val' Wed May 02 18:54:38 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gly12val,tyr40cys' Tue Jun 30 19:13:01 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.[Gly12Val;Tyr40Cys]' ecobb Escherichia coli (strain B / BL21) physical association physical association 2C 2D 3 Note that the authors refer to COS cells. It is assumed that these are Cos-1 cells. Following coexpression of HA-tagged Ras mutants with Epac2, Ras proteins were immunoprecipitated with HA antibody, and Epac2 was found to co-precipitate with oncogenic Ras mutants, but not with the closely related M-RasLeu-71 that has an activating mutation equivalent to H-RasLeu-61 (Fig. 2C). Interestingly, Kand N-Ras bound more effectively to Epac2 than did H-Ras.Binding to activated H-Ras proteins containing Ser35, Gly37, or Cys40 mutations in their effector-binding loop was diminished (Fig. 2D). Binding to activated H-Ras proteins containing Ser35, Gly37, or Cys40 mutations in their effector-binding loop as well as the Val12 mutation was diminished compared to the protein with only the Val12 mutation. Note it was shown that the deletion mutant Epac2DeltaRA did not coimmuniprecipitate with H-Ras12V. hras-rapgef4-2 3 5 unspecified role unspecified role neutral component neutral component n-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range 4 unspecified role unspecified role neutral component neutral component region GFP-H-RasLeu61 gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined P01112:p.Gln61Leu mutation mutation certain certain sequence position certain certain certain sequence position certain Q L Wed May 02 15:39:14 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'gln61leu' Tue Jun 30 15:56:40 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Gln61Leu' colocalization colocalization 4 C, D upon coexpression with GFP-H-RasLeu-61 (Fig. 4, C and D), the GEF was predominantly redistributed to the plasma membrane (Fig. 4D) where it colocalized with Ras (Fig. 4C).