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/ http://www.ebi.ac.uk/ cho-2005-2 Raf-1 is a binding partner of DSCR1. in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Cho YJ., Abe M., Kim SY., Sato Y. Cancer - Interactions investigated in the context of cancer A unique UniProtKB entry could not be associated with calcineurin since the exact gene product of the interactor cannot be ascertained, as it is a multi-subunit protein. y-sato@idac.tohoku.ac.jp Arch. Biochem. Biophys. (0003-9861) 2005 cho-2005-1 Raf-1 is a binding partner of DSCR1. in vitro In vitro pull down pull down ms participant Identification by mass spectrometry ms participant Cho YJ., Abe M., Kim SY., Sato Y. Cancer - Interactions investigated in the context of cancer A unique UniProtKB entry could not be associated with calcineurin since the exact gene product of the interactor cannot be ascertained, as it is a multi-subunit protein. y-sato@idac.tohoku.ac.jp Arch. Biochem. Biophys. (0003-9861) 2005 cho-2005-3 Raf-1 is a binding partner of DSCR1. bovin-gm_7373 Bos taurus endothelial aorta cell line gm_7373 Bovine endothelial aorta cell line Bovine endothelial aorta established from a male bovine fetus at 24-week gestation; vascular endothelium transformed with benzo(a)pyrene in vitro. anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Cho YJ., Abe M., Kim SY., Sato Y. Cancer - Interactions investigated in the context of cancer A unique UniProtKB entry could not be associated with calcineurin since the exact gene product of the interactor cannot be ascertained, as it is a multi-subunit protein. y-sato@idac.tohoku.ac.jp Arch. Biochem. Biophys. (0003-9861) 2005 cho-2005-4 Raf-1 is a binding partner of DSCR1. in vitro In vitro pull down pull down weight by comassie molecular weight estimation by coomasie staining weight by comassie Cho YJ., Abe M., Kim SY., Sato Y. Cancer - Interactions investigated in the context of cancer A unique UniProtKB entry could not be associated with calcineurin since the exact gene product of the interactor cannot be ascertained, as it is a multi-subunit protein. y-sato@idac.tohoku.ac.jp Arch. Biochem. Biophys. (0003-9861) 2005 cho-2005-5 Raf-1 is a binding partner of DSCR1. in vitro In vitro enzymatic study enzymatic study weight silver stain molecular weight estimation by silver staining weight silver stain Cho YJ., Abe M., Kim SY., Sato Y. Cancer - Interactions investigated in the context of cancer A unique UniProtKB entry could not be associated with calcineurin since the exact gene product of the interactor cannot be ascertained, as it is a multi-subunit protein. GST-DSCR1.4 was incubated with mu-calpain from human erythrocytes. Reactions terminated by adding SDS sample buffer. Samples were analyzed by SDS-PAGE and silver staining. A unique UniProtKB entry could not be associated with calcineurin since the exact gene product of the interactor cannot be ascertained, as it is a multi-subunit protein. y-sato@idac.tohoku.ac.jp Arch. Biochem. Biophys. (0003-9861) 2005 p53805-2 Calcipressin-1 RCAN1 ADAPT78 CSP1 DSC1 DSCR1 Adapt78 Down syndrome critical region protein 1 Myocyte-enriched calcineurin-interacting protein 1 Regulator of calcineurin 1 CALP1-S protein protein human Homo sapiens Human MHFRNFNYSFSSLIACVANSDIFSESETRAKFESLFRTYDKDITFQYFKSFKRVRINFSNPFSAADARLQLHKTEFLGKEMKLYFAQTLHIGSSHLAPPNPDKQFLISPPASPPVGWKQVEDATPVINYDLLYAISKLGPGEKYELHAATDTTPSVVVHVCESDQEKEEEEEMERMRRPKPKIIQTRRPEYTPIHLS raf1_human RAF proto-oncogene serine/threonine-protein kinase RAF1 RAF Proto-oncogene c-RAF Raf-1 protein protein human Homo sapiens Human MEHIQGAWKTISNGFGFKDAVFDGSSCISPTIVQQFGYQRRASDDGKLTDPSKTSNTIRVFLPNKQRTVVNVRNGMSLHDCLMKALKVRGLQPECCAVFRLLHEHKGKKARLDWNTDAASLIGEELQVDFLDHVPLTTHNFARKTFLKLAFCDICQKFLLNGFRCQTCGYKFHEHCSTKVPTMCVDWSNIRQLLLFPNSTIGDSGVPALPSLTMRRMRESVSRMPVSSQHRYSTPHAFTFNTSSPSSEGSLSQRQRSTSTPNVHMVSTTLPVDSRMIEDAIRSHSESASPSALSSSPNNLSPTGWSQPKTPVPAQRERAPVSGTQEKNKIRPRGQRDSSYYWEIEASEVMLSTRIGSGSFGTVYKGKWHGDVAVKILKVVDPTPEQFQAFRNEVAVLRKTRHVNILLFMGYMTKDNLAIVTQWCEGSSLYKHLHVQETKFQMFQLIDIARQTAQGMDYLHAKNIIHRDMKSNNIFLHEGLTVKIGDFGLATVKSRWSGSQQVEQPTGSVLWMAPEVIRMQDNNPFSFQSDVYSYGIVLYELMTGELPYSHINNRDQIIFMVGRGYASPDLSKLYKNCPKAMKRLVADCVKKVKEERPLFPQILSSIELLQHSLPKINRSASEPSLHRAAHTEDINACTLTTSPRLPVF raf1_bovin RAF proto-oncogene serine/threonine-protein kinase RAF1 Proto-oncogene c-RAF Raf-1 protein protein bovin Bos taurus (Bovine) Bovine MEHIQGAWKTISNGFGFKDTVFDGTSCISPTIVQQFGYQRRASDDGKLTDPSKTSNTIRVFLPNKQRTVVNVRNGMSLHDCLMKALKVRGLQPECCAVFRLLHEHKGKKARLDWNTDAASLIGEELQVDFLDHVPLTTHNFARKTFLKLAFCDICQKFLLNGFRCQTCGYKFHEHCSTKVPTMCVDWSNIRQLLLFPNSTVGDGGVPALPSLTMRRMRESVSRIPPGSQHRYSTPHAFTFSASSPSSEGSLSQRQRSTSTPNVHMVSATLPVDSRMIEDAIRSHSESGSPSALSSSPNNLSPTGWSQPKTPAPAQRERAPGSSTQEKNKIRPRGQRDSSYYWEIEASEVMLSTRIGSGSFGTVYKGKWHGDVAVKILKVVDPTPEQFQAFRNEVAVLRKTRHVNILLFMGYMTKDNLAIVTQWCEGSSLYKHLHVQETKFQMFQLIDIARQTAQGMDYLHAKNIIHRDMKSNNIFLHEGLTVKIGDFGLATVKSRWSGSQQVEQPTGSILWMAPEVIRMQDNNPFSFQSDVYSYGIVLYELMTGELPYSHINNRDQIIFMVGRGYASPDLSKLYKNCPKAMKRLVADCVKKVKEERPLFPQILSSIELLQHSLPKINRSASEPSLHRAAHTEDINACTLTTSPRLPVF can1_human Calpain-1 catalytic subunit CAPN1 CANPL1 Calpain-1 large subunit Calcium-activated neutral proteinase 1 Calpain mu-type Micromolar-calpain Cell proliferation-inducing gene 30 protein PIG30 protein protein human Homo sapiens Human MSEEIITPVYCTGVSAQVQKQRARELGLGRHENAIKYLGQDYEQLRVRCLQSGTLFRDEAFPPVPQSLGYKDLGPNSSKTYGIKWKRPTELLSNPQFIVDGATRTDICQGALGDCWLLAAIASLTLNDTLLHRVVPHGQSFQNGYAGIFHFQLWQFGEWVDVVVDDLLPIKDGKLVFVHSAEGNEFWSALLEKAYAKVNGSYEALSGGSTSEGFEDFTGGVTEWYELRKAPSDLYQIILKALERGSLLGCSIDISSVLDMEAITFKKLVKGHAYSVTGAKQVNYRGQVVSLIRMRNPWGEVEWTGAWSDSSSEWNNVDPYERDQLRVKMEDGEFWMSFRDFMREFTRLEICNLTPDALKSRTIRKWNTTLYEGTWRRGSTAGGCRNYPATFWVNPQFKIRLDETDDPDDYGDRESGCSFVLALMQKHRRRERRFGRDMETIGFAVYEVPPELVGQPAVHLKRDFFLANASRARSEQFINLREVSTRFRLPPGEYVVVPSTFEPNKEGDFVLRFFSEKSAGTVELDDQIQANLPDEQVLSEEEIDENFKALFRQLAGEDMEISVKELRTILNRIISKHKDLRTKGFSLESCRSMVNLMDRDGNGKLGLVEFNILWNRIRNYLSIFRKFDLDKSGSMSAYEMRMAIESAGFKLNKKLYELIITRYSEPDLAVDFDNFVCCLVRLETMFRFFKTLDTDLDGVVTFDLFKWLQLTMFA rcan1-raf1-1 GST-DSCR1.4 pulls down Raf-1 from GM7373 lysates 670721 N/A 670726 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 false ecoli Escherichia coli (strain K12) N/A 670727 unspecified role unspecified role prey prey bovin Bos taurus (Bovine) Bovine physical association physical association false false false 1 B rcan1-raf1-2 GST-DSCR1.4 pulls down Raf-1 from MS-1 endothelial cell lysates 670721 N/A 670727 unspecified role unspecified role prey prey mouse-ms_1 Mus musculus pancreatic islet endothelial cell line ms_1 MS-1 (MILE SVEN 1) murine pancreatic islet endothelial cell line N/A 670726 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 false ecoli Escherichia coli (strain K12) physical association physical association false false false 1 B rcan1-raf1 GST-DSCR1.4 pulls down Raf-1 from HUVEC lysates 670722 N/A 670726 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 false ecoli Escherichia coli (strain K12) N/A 670727 unspecified role unspecified role prey prey human-uvec Human Umbilical Vein Endothelial Cells (HUVEC) huvec umbilical_vein_endothelium Umbilical vein endothelial cell [Umbilical vein endothelium] physical association physical association false false false 1 A rcan1-raf1-3 GFP-DSCR1.4 coimmunoprecipitates RAF1 from GM7373 lysates 670723 N/A 670728 unspecified role unspecified role prey prey N/A 670726 unspecified role unspecified role bait bait n-terminal gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false n-terminal sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false c-terminal sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) physical association physical association false false false 1C, 4 The first six amino acids of DSCR1.4 are necessary for binding of the N-terminal fragment as GST-DSCR1.4 aa 7-133 did not bind RAF-1. Thus it seems cleavage between Phe-6 and Asn-7 abolishes Raf-1 binding through the N-terminal of DSCR1.4. It also seems cleavage between Tyr-133 and Ala-134 releases C-terminal Raf-1 binding domain from the inhibition by aa 115-133. rcan1-raf1-4 GST-DSCR1.4 (aa 1-84) and GST-DSCR1.4 (aa 143-197) are sufficient to pull down Raf-1 from HUVEC lysates 670724 N/A 670727 unspecified role unspecified role prey prey human-uvec Human Umbilical Vein Endothelial Cells (HUVEC) huvec umbilical_vein_endothelium Umbilical vein endothelial cell [Umbilical vein endothelium] N/A 670726 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 false n-terminal sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false c-terminal sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) physical association physical association false false false 2 The results suggest that Raf-1 can associate with two regions of DSCR1.4: one in the N-terminal end (amino acids 1-84) and the other in the C-terminus (amino acids 134-156), and this is interfered with by amino acid sequence 115-133 of DSCR1.4. capn1-rcan1 Mu-calpain cleaves DSCR1.4 in vitro 670725 N/A 670726 enzyme target enzyme target substrate substrate unspecified role unspecified role 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 false ecoli Escherichia coli (strain K12) N/A 670729 enzyme enzyme unspecified role unspecified role human-erythrocyte Homo sapiens erythrocyte Erythrocyte, Red blood cell; Corpuscle; RBC protein cleavage protein cleavage false false false 3 GST-DSCR1.4 was cleaved into multiple fragments of a molecular mass that ranged from 40 to 12 kDa. Two of the cleavage sites are between phe-6 and asn-7 and between tyr-133 and ala-134.