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/ antoine_bertrand-2011-1 in vitro In vitro pull down pull down sequence tag sequence tag identification sequence tag Antoine-Bertrand J., Ghogha A., Luangrath V., Bedford FK., Lamarche-Vane N. At the time of curation supplementary materials were not available. From pmid 8416983 and 9230083: GST-ezrin, and GST-ezrinNT pCB6-ezrin-VSVG, pCB6-ezrinT567D-VSVG, pCB6-ezrinT567A-VSVG, pCB6-ezrinΔ29-VSVG) are all of human origin. From pmid: 15557120 All DCC construct are from rat. According to IMEx rules, Interactions detected with aspecific antibodies (e.g. anti-pERM) were not captured. nathalie.lamarche@mcgill.ca Molecular biology of the cell 2011 imex curation Only protein-protein interactions imex curation antoine_bertrand-2011-2 in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Antoine-Bertrand J., Ghogha A., Luangrath V., Bedford FK., Lamarche-Vane N. At the time of curation supplementary materials were not available. From pmid 8416983 and 9230083: GST-ezrin, and GST-ezrinNT pCB6-ezrin-VSVG, pCB6-ezrinT567D-VSVG, pCB6-ezrinT567A-VSVG, pCB6-ezrinΔ29-VSVG) are all of human origin. From pmid: 15557120 All DCC construct are from rat. According to IMEx rules, Interactions detected with aspecific antibodies (e.g. anti-pERM) were not captured. nathalie.lamarche@mcgill.ca Molecular biology of the cell 2011 imex curation Only protein-protein interactions imex curation antoine_bertrand-2011-3 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 bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Antoine-Bertrand J., Ghogha A., Luangrath V., Bedford FK., Lamarche-Vane N. At the time of curation supplementary materials were not available. From pmid 8416983 and 9230083: GST-ezrin, and GST-ezrinNT pCB6-ezrin-VSVG, pCB6-ezrinT567D-VSVG, pCB6-ezrinT567A-VSVG, pCB6-ezrinΔ29-VSVG) are all of human origin. From pmid: 15557120 All DCC construct are from rat. According to IMEx rules, Interactions detected with aspecific antibodies (e.g. anti-pERM) were not captured. nathalie.lamarche@mcgill.ca Molecular biology of the cell 2011 imex curation Only protein-protein interactions imex curation antoine_bertrand-2011-4 rat-embryonic_brain rat-embryonic_brain Rat embryonic_brain anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Antoine-Bertrand J., Ghogha A., Luangrath V., Bedford FK., Lamarche-Vane N. At the time of curation supplementary materials were not available. From pmid 8416983 and 9230083: GST-ezrin, and GST-ezrinNT pCB6-ezrin-VSVG, pCB6-ezrinT567D-VSVG, pCB6-ezrinT567A-VSVG, pCB6-ezrinΔ29-VSVG) are all of human origin. From pmid: 15557120 All DCC construct are from rat. According to IMEx rules, Interactions detected with aspecific antibodies (e.g. anti-pERM) were not captured. nathalie.lamarche@mcgill.ca Molecular biology of the cell 2011 imex curation Only protein-protein interactions imex curation antoine_bertrand-2011-5 mouse-n1e-115 Mus musculus neuroblastoma cells n1e-115 Mouse neuroblastoma anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Antoine-Bertrand J., Ghogha A., Luangrath V., Bedford FK., Lamarche-Vane N. At the time of curation supplementary materials were not available. From pmid 8416983 and 9230083: GST-ezrin, and GST-ezrinNT pCB6-ezrin-VSVG, pCB6-ezrinT567D-VSVG, pCB6-ezrinT567A-VSVG, pCB6-ezrinΔ29-VSVG) are all of human origin. From pmid: 15557120 All DCC construct are from rat. According to IMEx rules, Interactions detected with aspecific antibodies (e.g. anti-pERM) were not captured. nathalie.lamarche@mcgill.ca Molecular biology of the cell 2011 imex curation Only protein-protein interactions imex curation e9pt65_rat Rdx protein protein rat Rattus norvegicus (Rat) Rat MPKPINVRVTTMDAELEFAIQPNTTGKQLFDQVVKTVGLREVWFFGLQYVDSKGYSTWLKLNKKVTQQDVKKENPLQFKFRAKFFPEDVSEELIQEITQRLFFLQVKEAILNDEIYCPPETAVLLASYAVQAKYGDYNKEIHKPGYLANDRLLPQRVLEQHKLTKEQWEERIQNWHEEHRGMLREDSMMEYLKIAQDLEMYGVNYFEIKNKKGTELWLGVDALGLNIYEHDDNFVPSTLFPLTKEAKVAHCDEIFLIKMIDFQSDDFVFYAPRLRINKRILALCMGNHELYMRRRKPDTIEVQQMKAQAREEKHQKQLERAQLENEKKKREIAEKEKERIEREKEELMERLRQIEEQTMKAQKELEEQTRKALELEQERQRAKEEAERLDRERRAAEEAKSAIAKQAADQMKNQEQLAAELAEFTAKIALLEEAKKKKEEEATEWQHKAFAAQEDLEKTKEELKTVMCAPPPPPPPAVVPPTENEHDEQDEHSAEASAELSSEGVTNHRSEEERVTETQKNERVKKQLQALSSELAQARDETKKTQNDVLHAENVKAGRDKYKTLRQIRQGNTKQRIDEFEAM moes_rat Moesin Msn Membrane-organizing extension spike protein protein protein rat Rattus norvegicus (Rat) Rat MPKTISVRVTTMDAELEFAIQPNTTGKQLFDQVVKTIGLREVWFFGLQYQDTKAFSTWLKLNKKVTAQDVRKESPLLFKFRAKFYPEDVSEELIQDITQRLFFLQVKEGILNDDIYCPPETAVLLASYAVQSKYGDFNKEVHKSGYLAGDKLLPQRVLEQHKLNKDQWEERIQVWHEEHRGMLREDAVLEYLKIAQDLEMYGVNYFSIKNKKGSELWLGVDALGLNIYEQNDRLTPKIGFPWSEIRNISFNDKKFVIKPIDKKAPDFVFYAPRLRINKRILALCMGNHELYMRRRKPDTIEVQQMKAQAREEKHQKQMERALLENEKKKRELAEKEKEKIEREKEELMEKLKQIEEQTKKAQQELEEQTRRALELEQERKRAQSEAEKLAKERQEAEEAKEALLQASRDQKKTQEQLASEMAELTARVSQLEMARKKKESEAEECHQKAQMVQEDLEKTRAELKTAMSTPHVAEPAENEHDEQDENGAEASAELRADAMAKDRSEEERTTEAEKNERVQKHLKALTSELANARDESKKTTNDMIHAENMRLGRDKYKTLRQIRQGNTKQRIDEFESM ezri_rat Ezrin Vil2 p81 Cytovillin Villin-2 Ezr protein protein rat Rattus norvegicus (Rat) Rat MPKPINVRVTTMDAELEFAIQPNTTGKQLFDQVVKTIGLREVWYFGLQYVDNKGFPTWLKLDKKVSAQEVRKENPVQFKFRAKFYPEDVADELIQDITQKLFFLQVKEGILSDEIYCPPETAVLLGSYAVQAKFGDYNKEMHKSGYLSSERLIPQRVMDQHKLSRDQWEDRIQVWHAEHRGMLKDSAMLEYLKIAQDLEMYGINYFEIKNKKGTDLWLGVDALGLNIYEKDDKLTPKIGFPWSEIRNISFNDKKFVIKPIDKKAPDFVFYAPRLRINKRILQLCMGNHELYMRRRKPDTIEVQQMKAQAREEKHQKQLERQQLETEKKRRETVEREKEQMLREKEELMLRLQDFEQKTKRAEKELSEQIEKALQLEEERRRAQEEAERLEADRMAALRAKEELERQAQDQIKSQEQLAAELAEYTAKIALLEEARRRKEDEVEEWQHRAKEAQDDLVKTKEELHLVMTAPPPPPPPVYEPVNYHVQEGLQDEGAEPMGYSAELSSEGILDDRNEEKRITEAEKNERVQRQLLTLSNELSQARDENKRTHNDIIHNENMRQGRDKYKTLRQIRQGNTKQRIDEFEAM dcc_rat Netrin receptor DCC Dcc Tumor suppressor protein DCC protein protein rat Rattus norvegicus (Rat) Rat MENSLGCVWVPKLAFVLFGASLLSAHLQVTGFQIKPFTSLHFVSEPSDAVTMRGGNVLLNCSAESDRGVPVIKWKKDGLILALGMDDRKQQLPNGSLLIQNILHSRHHKPDEGLYQCEASLGDSGSIISRTAKVMVAGPLRFLSQTESITAFMGDTVLLKCEVIGDPMPTIHWQKNQQDLNPIPGDSRVVVLPSGALQISRLQPGDSGVYRCSARNPASTRTGNEAEVRILSDPGLHRQLYFLQRPSNVIAIEGKDAVLECCVSGYPPPSFTWLRGEEVIQLRSKKYSLLGGSNLLISNVTDDDSGTYTCVVTYKNENISASAELTVLVPPWFLNHPSNLYAYESMDIEFECAVSGKPVPTVNWMKNGDVVIPSDYFQIVGGSNLRILGVVKSDEGFYQCVAENEAGNAQSSAQLIVPKPAIPSSSILPSAPRDVVPVLVSSRFVRLSWRPPAEAKGNIQTFTVFFSREGDNRERALNTTQPGSLQLTVGNLKPEAMYTFRVVAYNEWGPGESSQPIKVATQPELQVPGPVENLHAVSASPTSILITWEPPAYANGPVQGYRLFCTEVSTGKEQNIEVDGLSYKLEGLKKFTEYTLRFLAYNRYGPGVSTDDITVVTLSDVPSAPPQNVSLEVVNSRSIKVSWLPPPSGTQNGFITGYKIRHRKTTRRGEMETLEPNNLWYLFTGLEKGSQYSFQVSAMTVNGTGPPSNWYTAETPENDLDESQVPDQPSSLHVRPQTNCIIMSWTPPLNPNIVVRGYIIGYGVGSPYAETVRVDSKQRYYSIERLESSSHYVISLKAFNNAGEGVPLYESATTRSITDPTDPVDYYPLLDDFPTSGPDVSTPMLPPVGVQAVALTHEAVRVSWADNSVPKNQKTSDVRLYTVRWRTSFSASAKYKSEDTTSLSYTATGLKPNTMYEFSVMVTKNRRSSTWSMTAHATTYEAAPTSAPKDLTVITREGKPRAVIVSWQPPLEANGKITAYILFYTLDKNIPIDDWIMETISGDRLTHQIMDLSLDTMYYFRIQARNAKGVGPLSDPILFRTLKVEHPDKMANDQGRHGDGGYWPVDTNLIDRSTLNEPPIGQMHPPHGSVTPQKNSNLLVITVVTVGVLTVLVVVIVAVICTRRSSAQQRKKRATHSASKRKGSQKDLRPPDLWIHHEEMEMKNIEKPAGTDPAGRGSPIQSCQDLTPVSHSQSESQMGSKSASHSGQDTEEAGSSMSTLERSLAARRATRTKLMIPMEAQSNNPAVVSAIPVPTLESAQYPGILPSPTCGYPHPQFTLRPVPFPTLSVDRGFGAGRTVSEGPTAQQQPMLPPAQPEHPSSEEAPSRTIPTACVRPTHPLRSFANPLLPPPMSAIEPKVPYTPLLSQPGPTLPKTHVKTASLGLAGKARSPLLPVSVPTAPEVSEESHKPTEDPASVYEQDDLSEQMASLEGLMKQLNAITGSAF ezri_human Ezrin EZR VIL2 p81 Cytovillin Villin-2 protein protein human Homo sapiens Human MPKPINVRVTTMDAELEFAIQPNTTGKQLFDQVVKTIGLREVWYFGLHYVDNKGFPTWLKLDKKVSAQEVRKENPLQFKFRAKFYPEDVAEELIQDITQKLFFLQVKEGILSDEIYCPPETAVLLGSYAVQAKFGDYNKEVHKSGYLSSERLIPQRVMDQHKLTRDQWEDRIQVWHAEHRGMLKDNAMLEYLKIAQDLEMYGINYFEIKNKKGTDLWLGVDALGLNIYEKDDKLTPKIGFPWSEIRNISFNDKKFVIKPIDKKAPDFVFYAPRLRINKRILQLCMGNHELYMRRRKPDTIEVQQMKAQAREEKHQKQLERQQLETEKKRRETVEREKEQMMREKEELMLRLQDYEEKTKKAERELSEQIQRALQLEEERKRAQEEAERLEADRMAALRAKEELERQAVDQIKSQEQLAAELAEYTAKIALLEEARRRKEDEVEEWQHRAKEAQDDLVKTKEELHLVMTAPPPPPPPVYEPVSYHVQESLQDEGAEPTGYSAELSSEGIRDDRNEEKRITEAEKNERVQRQLLTLSSELSQARDENKRTHNDIIHNENMRQGRDKYKTLRQIRQGNTKQRIDEFEAL dccp-y1418-1 3541228 N/A 3541233 unspecified role unspecified role prey prey cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level rat-embryonic_brain rat-embryonic_brain Rat embryonic_brain N/A 3541234 unspecified role unspecified role prey prey cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level rat-embryonic_brain rat-embryonic_brain Rat embryonic_brain N/A 3541235 unspecified role unspecified role prey prey cell lysate cell lysate physiological level physiological level endogenous level endogenous endogenous endogenous level rat-embryonic_brain rat-embryonic_brain Rat embryonic_brain N/A 3541236 weight by comassie molecular weight estimation by coomasie staining weight by comassie unspecified role unspecified role bait bait purified purified region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false phosphorylated Tyr optyr O4'-phospho-L-tyrosine (2S)-2-amino-3-(4-phosphonooxyphenyl)propanoic acid 2-azanyl-3-(4-phosphonooxyphenyl)propanoic acid MOD_RES Phosphotyrosine O4-phosphotyrosine tyrosine phosphate 2-amino-3-(4-hydroxyphenyl)propanoic acid 4'-phosphate O4'-phospho-L-tyrosine O4'-phosphorylated L-tyrosine OPTyr Phospho Phospho Tyrosinyl Phosphorylation phosphorylationy certain certain sequence position certain certain certain sequence position certain false chemical synthesis Chemical synthesis (Chemical synthesis) association association false false false Fig. 1A ezrin-dcc-1 3541229 N/A 3541236 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed cell lysate cell lysate 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. N/A 3541237 unspecified role unspecified role bait bait purified purified gst tag 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 region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) physical association physical association false false false Fig. 1C ezrin-dcc-2 3541230 N/A 3541236 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain false N/A 3541237 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain false vsvg tag vsv tag vsv tag vesicular stomatitis virus tag vesicular stomatitis virus tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false physical association physical association false false false Fig. 2B When ezrin was coexpressed with DCC in HEK293 cells stimulated with netrin-1, we observed that ezrin was able to interact with DCC in a netrin-1–dependent manner ( Figure 2B). However, DCC lacking the intracellular domain (DCC (1–1120)) or inactive ezrin lacking the F-actin binding domain (ezrinΔ29) abolished the ezrin–DCC interaction ( Figure 2B). netrin - Q90922 dcc-ezr-1 3541231 N/A 3541236 unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level N/A 3541235 unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level physical association physical association false false false Fig. 2C In rat E18 cortical neurons at 2DIV, netrin-1 transiently stimulated the interaction of ezrin and DCC ( Figure 2E). Coimmunoprecipitation of ezrin with DCC was observed after 5 min of netrin-1 stimulation and was significantly reduced after a 20-min stimulation ( Figure 2, E and F). dcc-ezr-2 3541232 N/A 3541236 unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell N/A 3541237 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell vsvg tag vsv tag vsv tag vesicular stomatitis virus tag vesicular stomatitis virus tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false P15311:p.Thr567Asp mutation mutation certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Fig. 2A