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 adaptor Grb7 is a novel calmodulin-binding protein: functional implications of the interaction of calmodulin with Grb7. The adaptor Grb7 is a novel calmodulin-binding protein: functional implications of the interaction of calmodulin with Grb7. Oncogene (0950-9232) 2005 imex curation Li H., Sanchez-Torres J., del Carpio AF., Nogales-Gonzalez A., Molina-Ortiz P., Moreno MJ., Torok K., Villalobo A. Origin of P62157 taken from manufacturer's website antonio.villalobo@iib.uam.es 29-OCT-2007: Contacted by JYOTI. 2007-06-01: Antonio Villalobo, Instituto de Investigaciones Biomedicas, Universidad Autonoma de Madrid, Spain Only protein-protein interactions in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Li H., Sanchez-Torres J., del Carpio AF., Nogales-Gonzalez A., Molina-Ortiz P., Moreno MJ., Torok K., Villalobo A. Origin of P62157 taken from manufacturer's website antonio.villalobo@iib.uam.es Accepted 2007-JUL-14 by JYOTI. 2007-06-01: Antonio Villalobo, Instituto de Investigaciones Biomedicas, Universidad Autonoma de Madrid, Spain Oncogene (0950-9232) 2005 Only protein-protein interactions imex curation The adaptor Grb7 is a novel calmodulin-binding protein: functional implications of the interaction of calmodulin with Grb7. The adaptor Grb7 is a novel calmodulin-binding protein: functional implications of the interaction of calmodulin with Grb7. Oncogene (0950-9232) 2005 imex curation Li H., Sanchez-Torres J., del Carpio AF., Nogales-Gonzalez A., Molina-Ortiz P., Moreno MJ., Torok K., Villalobo A. Origin of P62157 taken from manufacturer's website antonio.villalobo@iib.uam.es 29-OCT-2007: Contacted by JYOTI. 2007-06-01: Antonio Villalobo, Instituto de Investigaciones Biomedicas, Universidad Autonoma de Madrid, Spain Only protein-protein interactions in vitro In vitro far western blotting far western blotting Affinity blotting Affinity blotting western blot western blot Immuno blot Immuno blot Li H., Sanchez-Torres J., del Carpio AF., Nogales-Gonzalez A., Molina-Ortiz P., Moreno MJ., Torok K., Villalobo A. Origin of P62157 taken from manufacturer's website antonio.villalobo@iib.uam.es Accepted 2007-JUL-14 by JYOTI. 29-OCT-2007: Contacted by JYOTI. 2007-06-01: Antonio Villalobo, Instituto de Investigaciones Biomedicas, Universidad Autonoma de Madrid, Spain Oncogene (0950-9232) 2005 Only protein-protein interactions imex curation The adaptor Grb7 is a novel calmodulin-binding protein: functional implications of the interaction of calmodulin with Grb7. The adaptor Grb7 is a novel calmodulin-binding protein: functional implications of the interaction of calmodulin with Grb7. Oncogene (0950-9232) 2005 imex curation Li H., Sanchez-Torres J., del Carpio AF., Nogales-Gonzalez A., Molina-Ortiz P., Moreno MJ., Torok K., Villalobo A. Origin of P62157 taken from manufacturer's website antonio.villalobo@iib.uam.es 29-OCT-2007: Contacted by JYOTI. 2007-06-01: Antonio Villalobo, Instituto de Investigaciones Biomedicas, Universidad Autonoma de Madrid, Spain Only protein-protein interactions in vitro In vitro filter binding filter binding Filter overlay assay Filter overlay assay western blot western blot Immuno blot Immuno blot Li H., Sanchez-Torres J., del Carpio AF., Nogales-Gonzalez A., Molina-Ortiz P., Moreno MJ., Torok K., Villalobo A. Immobilized phosphoinositides provided by the PIP Strips system Origin of P62157 taken from manufacturer's website antonio.villalobo@iib.uam.es Accepted 2007-JUL-14 by JYOTI. 29-OCT-2007: Contacted by JYOTI. 2007-06-01: Antonio Villalobo, Instituto de Investigaciones Biomedicas, Universidad Autonoma de Madrid, Spain Oncogene (0950-9232) 2005 Only protein-protein interactions imex curation 1-phosphatidyl-1d-myo-inositol 3,4,5-trisphosphate 1-phosphatidyl-1D-myo-inositol 3,4,5-trisphosphates small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) calm_rat Calmodulin Calm1 Calm Cam Cam1 Calm2 Cam2 Camb Calm3 Cam3 Camc CaMII protein protein rat Rattus norvegicus (Rat) Rat MADQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEAELQDMINEVDADGNGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK DIP protein P62161 original sequence version: 125 DIP protein P62161 original sequence version: 51 DIP protein P62161 original sequence version: 55 DIP protein P62161 original sequence version: 59 DIP protein P62161 original sequence version: 80 DIP protein P62161 original sequence version: 81 DIP protein P62161 original sequence version: 132 DIP protein P62161 original sequence version: 96 6B4BC3FCDE10727B grb7_human Growth factor receptor-bound protein 7 GRB7 GRB7 adapter protein Epidermal growth factor receptor GRB-7 B47 protein protein human Homo sapiens Human MELDLSPPHLSSSPEDLCPAPGTPPGTPRPPDTPLPEEVKRSQPLLIPTTGRKLREEERRATSLPSIPNPFPELCSPPSQSPILGGPSSARGLLPRDASRPHVVKVYSEDGACRSVEVAAGATARHVCEMLVQRAHALSDETWGLVECHPHLALERGLEDHESVVEVQAAWPVGGDSRFVFRKNFAKYELFKSSPHSLFPEKMVSSCLDAHTGISHEDLIQNFLNAGSFPEIQGFLQLRGSGRKLWKRFFCFLRRSGLYYSTKGTSKDPRHLQYVADVNESNVYVVTQGRKLYGMPTDFGFCVKPNKLRNGHKGLRIFCSEDEQSRTCWLAAFRLFKYGVQLYKNYQQAQSRHLHPSCLGSPPLRSASDNTLVAMDFSGHAGRVIENPREALSVALEEAQAWRKKTNHRLSLPMPASGTSLSAAIHRTQLWFHGRISREESQRLIGQQGLVDGLFLVRESQRNPQGFVLSLCHLQKVKHYLILPSEEEGRLYFSMDDGQTRFTDLLQLVEFHQLNRGILPCLLRHCCTRVAL A6867C20AFD46F74 1-phosphatidyl-1d-myo-inositol 3,5-bisphosphate 1-phosphatidyl-1D-myo-inositol 3,5-bisphosphates small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) 1-phosphatidyl-1d-myo-inositol 3,4-bisphosphate 1-phosphatidyl-1D-myo-inositol 3,4-bisphosphate small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) 1-phosphatidyl-1d-myo-inositol 4,5-bisphosphate 1-phosphatidyl-1D-myo-inositol 4,5-bisphosphates small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) pi5p 1-phosphatidyl-1D-myo-inositol 5-phosphate small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) phosphatidic acids phosphatidic acids small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) calm_bovin Calmodulin CALM CAM protein protein bovin Bos taurus (Bovine) Bovine MADQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEAELQDMINEVDADGNGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK DIP protein P62157 original sequence version: 125 DIP protein P62157 original sequence version: 58 DIP protein P62157 original sequence version: null 6B4BC3FCDE10727B pi3p 1-phosphatidyl-1D-myo-inositol 3-phosphates small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) calm-grb7-1 1 6 unspecified role unspecified role prey prey human-skbr3 Homo sapiens SKBR3 skbr3 SKBR3 Species: human; Tissue: breast; Tumor: adenocarcinoma; Derived from: metastatic site: pleural effusion 12 unspecified role unspecified role bait bait bovin-brain Bos taurus brain brain Brain [cerebrum] physical association physical association Figure 1a shows that Grb7 can be isolated from the SK-BR-3 cytosolic fraction (CF) by Ca2+-dependent CaM-affinity chromatography upon EGTA ([ethylene bis(oxyethylenenitrilo)]-tetraacetic acid) elution (EGTA, fractions 1-3), while it is absent in the effluent before EGTA addition (Ca2+). Figure 1a Interaction is calcium dependent. calm-grb7-2 1 12 unspecified role unspecified role bait bait bovin-brain Bos taurus brain brain Brain [cerebrum] 6 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) physical association physical association A CE (7.7 mg protein) from E. coli expressing recombinant Grb7 was subjected to CaM-affinity chromatography and processed as indicated above (top panel). Figure 1b calm-grb7-3 2 5 unspecified role unspecified role bait bait region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role prey prey human-skbr3 Homo sapiens SKBR3 skbr3 SKBR3 Species: human; Tissue: breast; Tumor: adenocarcinoma; Derived from: metastatic site: pleural effusion physical association physical association Figure 2a shows that when proteins from SK-BR-3 cells extract were overlaid with biotinylated CaM, a specific signal was detected in the presence but not in the absence of Ca2+ (presence of EGTA) (right panel). Figure 2b Interaction is calcium dependent calm-grb7-4 2 6 unspecified role unspecified role prey prey region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 5 unspecified role unspecified role bait bait region biotin tag biotin tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) physical association physical association Moreover, Figure 2c (top panel) shows that recombinant Grb7 interacts with biotinylated CaM in the presence but not in the absence of Ca2+ (presence of EGTA). Figures 2 & 4 Interaction is calcium dependent grb7-pi3p-1 3 5 competitor competitor competitor prey prey ecoli Escherichia coli (strain K12) 13 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait region binding region binding-associated region binding region deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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. association association Figure 7b demonstrates that, in the presence of EGTA, wild-type Grb7 binds to immobilized phosphoinositides (center subpanel), and that Grb7Delta has lower phosphoinositide binding capacity, although it significantly retains the ability to associate to phosphatidyl-3-phosphate, and phosphatidyl-3,5-bisphosphate (right subpanel). Fig 7 Calmodulin grb7-pi4p-1 3 10 unspecified role unspecified role prey prey 5 competitor competitor competitor prey prey ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role bait bait region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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. association association Figure 7b demonstrates that, in the presence of EGTA, wild-type Grb7 binds to immobilized phosphoinositides (center subpanel), and that Grb7Delta has lower phosphoinositide binding capacity, although it significantly retains the ability to associate to phosphatidyl-3-phosphate, and phosphatidyl-3,5-bisphosphate (right subpanel). Fig 7 Calmodulin grb7-pi5p-1 3 5 competitor competitor competitor prey prey ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role bait bait region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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. 10 unspecified role unspecified role prey prey association association Figure 7b demonstrates that, in the presence of EGTA, wild-type Grb7 binds to immobilized phosphoinositides (center subpanel), and that Grb7Delta has lower phosphoinositide binding capacity, although it significantly retains the ability to associate to phosphatidyl-3-phosphate, and phosphatidyl-3,5-bisphosphate (right subpanel). Fig 7 Calmodulin grb7-pi34p2-1 3 8 unspecified role unspecified role prey prey 5 competitor competitor competitor prey prey ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role bait bait region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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. association association Figure 7b demonstrates that, in the presence of EGTA, wild-type Grb7 binds to immobilized phosphoinositides (center subpanel), and that Grb7Delta has lower phosphoinositide binding capacity, although it significantly retains the ability to associate to phosphatidyl-3-phosphate, and phosphatidyl-3,5-bisphosphate (right subpanel). Fig 7 Calmodulin grb7-pi35p2-1 3 5 competitor competitor competitor prey prey ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait region binding region binding-associated region binding region deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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. association association Figure 7b demonstrates that, in the presence of EGTA, wild-type Grb7 binds to immobilized phosphoinositides (center subpanel), and that Grb7Delta has lower phosphoinositide binding capacity, although it significantly retains the ability to associate to phosphatidyl-3-phosphate, and phosphatidyl-3,5-bisphosphate (right subpanel). Fig 7 Calmodulin grb7-pi45p2-1 3 5 competitor competitor competitor prey prey 6 unspecified role unspecified role bait bait region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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. 9 unspecified role unspecified role prey prey association association Figure 7b demonstrates that, in the presence of EGTA, wild-type Grb7 binds to immobilized phosphoinositides (center subpanel), and that Grb7Delta has lower phosphoinositide binding capacity, although it significantly retains the ability to associate to phosphatidyl-3-phosphate, and phosphatidyl-3,5-bisphosphate (right subpanel). Fig 7 Calmodulin grb7-pi345p2-1 3 5 inhibitor inhibitor prey prey ecoli Escherichia coli (strain K12) 4 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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. association association Figure 7b demonstrates that, in the presence of EGTA, wild-type Grb7 binds to immobilized phosphoinositides (center subpanel), and that Grb7Delta has lower phosphoinositide binding capacity, although it significantly retains the ability to associate to phosphatidyl-3-phosphate, and phosphatidyl-3,5-bisphosphate (right subpanel). Fig 7 Calmodulin grb7-pa-1 3 6 unspecified role unspecified role bait bait region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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. 11 unspecified role unspecified role prey prey direct interaction direct interaction Figure 7b demonstrates that, in the presence of EGTA, wild-type Grb7 binds to immobilized phosphoinositides (center subpanel), and that Grb7Delta has lower phosphoinositide binding capacity, although it significantly retains the ability to associate to phosphatidyl-3-phosphate, and phosphatidyl-3,5-bisphosphate (right subpanel). Fig 7 grb7-pi3p-2 3 13 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait region binding region binding-associated region binding region deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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. direct interaction direct interaction Figure 7b demonstrates that, in the presence of EGTA, wild-type Grb7 binds to immobilized phosphoinositides (center subpanel), and that Grb7Delta has lower phosphoinositide binding capacity, although it significantly retains the ability to associate to phosphatidyl-3-phosphate, and phosphatidyl-3,5-bisphosphate (right subpanel). Fig 7 grb7-pi4p-2 3 10 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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. direct interaction direct interaction Figure 7b demonstrates that, in the presence of EGTA, wild-type Grb7 binds to immobilized phosphoinositides (center subpanel), and that Grb7Delta has lower phosphoinositide binding capacity, although it significantly retains the ability to associate to phosphatidyl-3-phosphate, and phosphatidyl-3,5-bisphosphate (right subpanel). Fig 7 grb7-pi5p-2 3 6 unspecified role unspecified role bait bait region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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. 10 unspecified role unspecified role prey prey direct interaction direct interaction Figure 7b demonstrates that, in the presence of EGTA, wild-type Grb7 binds to immobilized phosphoinositides (center subpanel), and that Grb7Delta has lower phosphoinositide binding capacity, although it significantly retains the ability to associate to phosphatidyl-3-phosphate, and phosphatidyl-3,5-bisphosphate (right subpanel). Fig 7 grb7-pi34p2-2 3 8 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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. direct interaction direct interaction Figure 7b demonstrates that, in the presence of EGTA, wild-type Grb7 binds to immobilized phosphoinositides (center subpanel), and that Grb7Delta has lower phosphoinositide binding capacity, although it significantly retains the ability to associate to phosphatidyl-3-phosphate, and phosphatidyl-3,5-bisphosphate (right subpanel). Fig 7 grb7-pi35p2-2 3 7 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait region binding region binding-associated region binding region deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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. direct interaction direct interaction Figure 7b demonstrates that, in the presence of EGTA, wild-type Grb7 binds to immobilized phosphoinositides (center subpanel), and that Grb7Delta has lower phosphoinositide binding capacity, although it significantly retains the ability to associate to phosphatidyl-3-phosphate, and phosphatidyl-3,5-bisphosphate (right subpanel). Fig 7 grb7-pi45p2-2 3 6 unspecified role unspecified role bait bait region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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. 9 unspecified role unspecified role prey prey direct interaction direct interaction Figure 7b demonstrates that, in the presence of EGTA, wild-type Grb7 binds to immobilized phosphoinositides (center subpanel), and that Grb7Delta has lower phosphoinositide binding capacity, although it significantly retains the ability to associate to phosphatidyl-3-phosphate, and phosphatidyl-3,5-bisphosphate (right subpanel). Fig 7 grb7-pi345p2-2 3 4 unspecified role unspecified role prey prey 6 unspecified role unspecified role bait bait region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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. direct interaction direct interaction Figure 7b demonstrates that, in the presence of EGTA, wild-type Grb7 binds to immobilized phosphoinositides (center subpanel), and that Grb7Delta has lower phosphoinositide binding capacity, although it significantly retains the ability to associate to phosphatidyl-3-phosphate, and phosphatidyl-3,5-bisphosphate (right subpanel). Fig 7