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/ garbett-2010-1 in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Garbett D., LaLonde DP., Bretscher A. Source of 6xHis-EPI64 construct gifted by Dr.Janet Ingraffea was found to be human (PMID: 22219378); source of beta-catenin was not provided anywhere in the reference and hence Figure 9c was not captured apb5@cornell.edu The Journal of cell biology 2010 Only protein-protein interactions imex curation imex curation garbett-2010-2 human-jeg3 Homo sapiens choriocarcinoma JEG3 jeg3 Human choriocarcinoma JEG3 confocal microscopy confocal microscopy immunostaining immunostaining Garbett D., LaLonde DP., Bretscher A. apb5@cornell.edu The Journal of cell biology 2010 Only protein-protein interactions imex curation imex curation ezri_human Ezrin EZR VIL2 p81 Cytovillin Villin-2 protein protein human Homo sapiens Human MPKPINVRVTTMDAELEFAIQPNTTGKQLFDQVVKTIGLREVWYFGLHYVDNKGFPTWLKLDKKVSAQEVRKENPLQFKFRAKFYPEDVAEELIQDITQKLFFLQVKEGILSDEIYCPPETAVLLGSYAVQAKFGDYNKEVHKSGYLSSERLIPQRVMDQHKLTRDQWEDRIQVWHAEHRGMLKDNAMLEYLKIAQDLEMYGINYFEIKNKKGTDLWLGVDALGLNIYEKDDKLTPKIGFPWSEIRNISFNDKKFVIKPIDKKAPDFVFYAPRLRINKRILQLCMGNHELYMRRRKPDTIEVQQMKAQAREEKHQKQLERQQLETEKKRRETVEREKEQMMREKEELMLRLQDYEEKTKKAERELSEQIQRALQLEEERKRAQEEAERLEADRMAALRAKEELERQAVDQIKSQEQLAAELAEYTAKIALLEEARRRKEDEVEEWQHRAKEAQDDLVKTKEELHLVMTAPPPPPPPVYEPVSYHVQESLQDEGAEPTGYSAELSSEGIRDDRNEEKRITEAEKNERVQRQLLTLSSELSQARDENKRTHNDIIHNENMRQGRDKYKTLRQIRQGNTKQRIDEFEAL nhrf1_human Na(+)/H(+) exchange regulatory cofactor NHE-RF1 SLC9A3R1 NHERF NHERF1 Ezrin-radixin-moesin-binding phosphoprotein 50 Regulatory cofactor of Na(+)/H(+) exchanger Sodium-hydrogen exchanger regulatory factor 1 Solute carrier family 9 isoform A3 regulatory factor 1 protein protein human Homo sapiens Human MSADAAAGAPLPRLCCLEKGPNGYGFHLHGEKGKLGQYIRLVEPGSPAEKAGLLAGDRLVEVNGENVEKETHQQVVSRIRAALNAVRLLVVDPETDEQLQKLGVQVREELLRAQEAPGQAEPPAAAEVQGAGNENEPREADKSHPEQRELRPRLCTMKKGPSGYGFNLHSDKSKPGQFIRSVDPDSPAEASGLRAQDRIVEVNGVCMEGKQHGDVVSAIRAGGDETKLLVVDRETDEFFKKCRVIPSQEHLNGPLPVPFTNGEIQKENSREALAEAALESPRPALVRSASSDTSEELNSQDSPPKQDSTAPSSTSSSDPILDFNISLAMAKERAHQKRSSKRAPQMDWSKKNELFSNL tb10a_human TBC1 domain family member 10A TBC1D10A EPI64 TBC1D10 Rab27A-GAP-alpha EBP50-PDX interactor of 64 kDa protein protein human Homo sapiens Human MAKSNGENGPRAPAAGESLSGTRESLAQGPDAATTDELSSLGSDSEANGFAERRIDKFGFIVGSQGAEGALEEVPLEVLRQRESKWLDMLNNWDKWMAKKHKKIRLRCQKGIPPSLRGRAWQYLSGGKVKLQQNPGKFDELDMSPGDPKWLDVIERDLHRQFPFHEMFVSRGGHGQQDLFRVLKAYTLYRPEEGYCQAQAPIAAVLLMHMPAEQAFWCLVQICEKYLPGYYSEKLEAIQLDGEILFSLLQKVSPVAHKHLSRQKIDPLLYMTEWFMCAFSRTLPWSSVLRVWDMFFCEGVKIIFRVGLVLLKHALGSPEKVKACQGQYETIERLRSLSPKIMQEAFLVQEVVELPVTERQIEREHLIQLRRWQETRGELQCRSPPRLHGAKAILDAEPGPRPALQPSPSIRLPLDAPLPGSKAKPKPPKQAQKEQRKQMKGRGQLEKPPAPNQAMVVAAAGDACPPQHVPPKDSAPKDSAPQDLAPQVSAHHRSQESLTSQESEDTYL mint ezr-slc9a3r1-1 3555593 N/A 3555595 unspecified role unspecified role bait bait purified purified ecoli Escherichia coli (strain K12) N/A 3555596 unspecified role unspecified role prey prey purified purified region mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range false O14745:p.Phe355Ala mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false O14745:p.Leu358delinsLeuAla mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false p.Phe355_Leu358del mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false O14745:p.[Met346Ala;Trp348Ala] mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false O14745:p.Phe355Arg mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false O14745:p.Leu358del mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false human-jeg3 Homo sapiens choriocarcinoma JEG3 jeg3 Human choriocarcinoma JEG3 direct interaction direct interaction false false false Figure 2b The EBP50 (SLC9A3R1)-ezrin interaction is critical for microvilli; Mutations in EBP50 (SLC9A3R1) - p.Phe355_Leu358del, p.[Met346Ala;Trp348Ala], p.Phe355Arg, p.Leu358del mutations abolish microvilli; p.Phe355Ala was not able to restore microvilli; p.Leu358delinsLeuAla was fully competent to form microvilli slc9a3r1-tbc1d10a-1 3555593 N/A 3555597 unspecified role unspecified role bait bait purified purified region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection N/A 3555596 unspecified role unspecified role prey prey purified purified O14745:p.[Ser280Ala;Ser302Ala] mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false O14745:p.[Ser280Asp;Ser302Asp] mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false O14745:p.[Ser162Ala;Ser339Ala;Ser340Ala] mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false O14745:p.[Ser162Asp;Ser339Asp;Ser340Asp] mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false direct interaction direct interaction false false false Figure 9a ezr-slc9a3r1-3 3555593 N/A 3555597 unspecified role unspecified role prey prey purified purified region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection N/A 3555595 unspecified role unspecified role bait bait purified purified ecoli Escherichia coli (strain K12) N/A 3555596 unspecified role unspecified role prey prey purified purified O14745:p.[Ser280Ala;Ser302Ala] mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false O14745:p.[Ser280Asp;Ser302Asp] mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false O14745:p.[Ser162Ala;Ser339Ala;Ser340Ala] mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false O14745:p.[Ser162Asp;Ser339Asp;Ser340Asp] mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false association association false false false Figure 9b ezr-slc9a3r1-2 3555594 N/A 3555595 unspecified role unspecified role neutral component neutral component physiological level physiological level endogenous level endogenous endogenous endogenous level fixed cell fixed cell N/A 3555596 unspecified role unspecified role neutral component neutral component over-expressed over expressed level over-expressed fixed cell fixed cell region 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 false O14745:p.Leu358delinsLeuAla mutation with no effect mutation with no effect mutation with no effect mutation not affecting interaction certain certain sequence position certain certain certain sequence position certain false O14745:p.Leu358del mutation disrupting strength mutation disrupting interaction strength mutation disrupting strength certain certain sequence position certain certain certain sequence position certain false O14745:p.[Ser280Ala;Ser302Ala] mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false O14745:p.[Ser280Asp;Ser302Asp] mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false O14745:p.[Ser162Ala;Ser339Ala;Ser340Ala] mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false O14745:p.[Ser162Asp;Ser339Asp;Ser340Asp] mutation decreasing strength mutation decreasing interaction strength mutation decreasing strength certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false EBP50 (SIC9A3R1) was rendered siRNA resistance by introduction of mutations without affecting the protein sequence colocalization colocalization false false false Figure 3b, 4b, 5b, 6b, 7b, 10b, S3b Mutations of EBP50 S162,339,340A and S162,339,340D were not captured as there are two substitution occurring at the same position and cannot be to fit in our feature