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/ ortiz-2004-2 Identification of HAX-1 as a protein that binds bile salt export protein and regulates its abundance in the apical membrane of Madin-Darby canine kidney cells. yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid predetermined predetermined participant predetermined Ortiz DF., Moseley J., Calderon G., Swift AL., Li S., Arias IM. BioCreative - Critical Assessment of Information Extraction systems in Biology Fig 1 daniel.ortiz@tufts.edu Rat liver cDNA library used. J. Biol. Chem. (0021-9258) 2004 Only protein-protein interactions imex curation ortiz-2004-4 Identification of HAX-1 as a protein that binds bile salt export protein and regulates its abundance in the apical membrane of Madin-Darby canine kidney cells. 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 Ortiz DF., Moseley J., Calderon G., Swift AL., Li S., Arias IM. BioCreative - Critical Assessment of Information Extraction systems in Biology Fig 1 daniel.ortiz@tufts.edu Rat liver cDNA library used. J. Biol. Chem. (0021-9258) 2004 Only protein-protein interactions imex curation ortiz-2004-5 Identification of HAX-1 as a protein that binds bile salt export protein and regulates its abundance in the apical membrane of Madin-Darby canine kidney cells. rat-liver rattus norvegicus liver Rat liver liver anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Ortiz DF., Moseley J., Calderon G., Swift AL., Li S., Arias IM. BioCreative - Critical Assessment of Information Extraction systems in Biology Canalicular membrane vesicles (CMV) prepared from rat liver were used. Fig 1 daniel.ortiz@tufts.edu Rat liver cDNA library used. J. Biol. Chem. (0021-9258) 2004 Only protein-protein interactions imex curation ortiz-2004-3 Identification of HAX-1 as a protein that binds bile salt export protein and regulates its abundance in the apical membrane of Madin-Darby canine kidney cells. in vitro In vitro pull down pull down predetermined predetermined participant predetermined Ortiz DF., Moseley J., Calderon G., Swift AL., Li S., Arias IM. BioCreative - Critical Assessment of Information Extraction systems in Biology Fig 1 daniel.ortiz@tufts.edu Rat liver cDNA library used. J. Biol. Chem. (0021-9258) 2004 Only protein-protein interactions imex curation ortiz-2004-1 Identification of HAX-1 as a protein that binds bile salt export protein and regulates its abundance in the apical membrane of Madin-Darby canine kidney cells. yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Ortiz DF., Moseley J., Calderon G., Swift AL., Li S., Arias IM. BioCreative - Critical Assessment of Information Extraction systems in Biology Fig 1 daniel.ortiz@tufts.edu Rat liver cDNA library used. J. Biol. Chem. (0021-9258) 2004 Only protein-protein interactions imex curation ortiz-2004-6 Identification of HAX-1 as a protein that binds bile salt export protein and regulates its abundance in the apical membrane of Madin-Darby canine kidney cells. canlf-mdck Canis familiaris (dog) kidney epithelial cells Canis lupus familiaris Dog Canis familiaris mdck Canine Cocker Spaniel kidney epithelial cell line fluorescence imaging fluorescence microscopy fluorescence imaging predetermined predetermined participant predetermined Ortiz DF., Moseley J., Calderon G., Swift AL., Li S., Arias IM. BioCreative - Critical Assessment of Information Extraction systems in Biology Fig 1 daniel.ortiz@tufts.edu Rat liver cDNA library used. J. Biol. Chem. (0021-9258) 2004 Only protein-protein interactions imex curation mdr3_rat Phosphatidylcholine translocator ABCB4 Pgp3 Pgy2 Abcb4 Mdr2 ATP-binding cassette sub-family B member 4 P-glycoprotein 2 P-glycoprotein 3 Multidrug resistance protein 2 Multidrug resistance protein 3 protein protein rat Rattus norvegicus (Rat) Rat MDLEAARNGTARRLDGDFELGSISNQSREKKKKVNLIGPLTLFRYSDWQDKLFMLLGTAMAIAHGSGLPLMMIVFGEMTDKFVDNAGNFSLPVNFSLSMLNPGRILEEEMTRYAYYYSGLGGGVLLAAYIQVSFWTLAAGRQIRKIRQKFFHAILRQEMGWFDIKGTTELNTRLTDDISKISEGIGDKVGMFFQAIATFFAGFIVGFIRGWKLTLVIMAITAILGLSTAVWAKILSTFSDKELAAYAKAGAVAEEALGAIRTVIAFGGQNKELERYQKHLENAKKIGIKKAISANISMGIAFLLIYASYALAFWYGSTLVISKEYTIGNAMTVFFSILIGAFSVGQAAPCIDAFPNARGAAYVIFDIIDNNPKIDSFSERGHKPDSIKGNLEFSDVHFSYPSRANIKILKGLNLKVKSGQTVALVGNSGCGKSTTVQLLQRLYDPTEGTISIDGQDIRNFNVRCLREFIGVVSQEPVLFSTTIAENIRYGRGNVTMDEIKKAVKEANAYDFIMKLPQKFDTLVGDRGAQLSGGQKQRIAIARALVRNPKILLLDEATSALDTESEAEVQAALDKAREGRTTIVIAHRLSTVRNADVIAGFEDGVIVEQGSHSELIKKEGIYFRLVNMQTSGSQILSEEFEVELSDEKAAGGVAPNGWKARIFRNSTKKSLKSSRAHQNRLDVETNELDANVPPVSFLKVLRLNKTEWPYFVVGTLCAIANGALQPAFSIILSEMIAIFGPGDDTVKQQKCNMFSLVFLGLGVHSFFTFFLQGFTFGKAGEILTTRLRSMAFKAMLRQDMSWFDDHKNSTGALSTRLATDAAQVQGATGTRLALIAQNTANLGTGIIISFIYGWQLTLLLLSVVPFIAVAGIVEMKMLAGNAKRDKKEMEAAGKIATEAIENIRTVVSLTQERKFESMYVEKLHGPYRNSVRKAHIYGITFSISQAFMYFSYAGCFRFGSYLIVNGHMRFKDVILVFSAIVLGAVALGHASSFAPDYAKAKLSAAYLFSLFERQPLIDSYSREGMWPDKFEGSVTFNEVVFNYPTRANVPVLQGLSLEVKKGQTLALVGSSGCGKSTVVQLLERFYDPMAGTVLLDGQEAKKLNVQWLRAQLGIVSQEPILFDCSIAKNIAYGDNSRVVSQDEIVRAAKEANIHPFIETLPQKYETRVGDKGTQLSGGQKQRIAIARALIRQPRVLLLDEATSALDTESEKVVQEALDKAREGRTCIVIAHRLSTIQNADLIVVIDNGKVKEHGTHQQLLAQKGIYFSMVNIQAGTQNL hax1_rat HCLS1-associated protein X-1 Hax1 Hs1bp1 HS1-associating protein X-1 HS1-binding protein 1 protein protein rat Rattus norvegicus (Rat) Rat MSVFDLFRGFFGFPGPRSHRDPFFGGMTRDDDDDEDDEEEEDSGAWGRESYAFDGFHPTEEFGFSFSPRGGMRFHGNFGFDDLVRDFNSIFSEMGAWTLPSHSPELPGPESETPGVRLREGQTLRDSMLKYPDSHQPRIFEGVLESHAKPESSKPAPDWGSQGPFHRLDDTWPVSPHSRAREDKDLDSQVSQEGLGPLLQPQPKSYFKSISVTKITKPDGTVEEHRTVVDSEGRRETTVTHQEAHDSSRSDPDPPRSSALDDPFSILDLLLGRWFRSR abcbb_rat Bile salt export pump Abcb11 Bsep Spgp ATP-binding cassette sub-family B member 11 Sister of P-glycoprotein protein protein rat Rattus norvegicus (Rat) Rat MSDSVILRSVKKFGEENHAFESDGSHNNDKKSRLQDKMKEGDIRVGFFELFRFSSSKDIWLMLMGGVCALLHGMAQPGILIIFGIMTDIFIKYDIERQELEIPGKACVNNTIVWINSSFHQNMTNGTVCGLVDIESEMIKFSGIYAGVGMTVLILGYFQIRLWVITGARQIRRMRKIYFRRIMRMEIGWFDCTSVGELNSRFADDIEKINDAIADQLAHFLQRMSTAMCGLLLGFYRGWKLTLVILAVSPLIGIGAAVIGLSIAKFTELELKAYAKAGSIADEVLSSIRTVAAFGGENKEVERYEKNLVFAQRWGIWKGMVMGFFTGYMWCLIFFCYALAFWYGSTLVLDEEEYTPGTLVQIFLCVILAAMNIGHASSCLEIFSTGCSAATNIFQTIDRQPVIDCMSGDGYKLDRIKGEIEFHNVTFHYPSRPDVKILDNLSMVIKPGETTALVGSSGAGKSTALQLIQRFYDPCEGMVTLDGHDIRSLNIRWLRDQIGIVEQEPVLFSTTIAENIRFGREDATMEDIVQAAKDANAYNFIMALPQQFDTLVGEGGGQMSGGQKQRVAIARALIRNPKILLLDMATSALDNESEARVQEALNKIQHGHTIISVAHRLSTVRAADVIIGFEHGVAVERGTHEELLERKGVYFMLVTLQSQGDNAHKETSIMGKDATEGGTLERTFSRGSYRDSLRASIRQRSKSQLSLLTHDPPLAVADHKSSYKDSKDNDVLVEEVEPAPVRRILKYNIPEWHYILVGSLSAAINGAVTPIYSLLFSQLLGTFSLLDKEQQRSEIHSMCLFFVILGCVSIFTQFLQGYTFAKSGELLTKRLRKFGFKAMLGQDIGWFDDLRNNPGVLTTRLATDASQVQGATGSQVGMMVNSFTNIIAALLIAFFFSWKLSLIITIFFPFLALSGAVQTKMLTGFASQDKQALEKAGQITSEALSNIRTVAGIGVEGRFIKAFEVELQTSYKTAVRKANIYGLCFAFSQGIAFLANSAAYRYGGYLIAYEGLGFSHVFRVVSSVALSATAVGRTFSYTPSYAKAKISAARFFQLLDRKPPINVYSEAGEKWDNFQGKIDFIDCKFTYPSRPDIQVLNGLSVSVNPGQTLAFVGSSGCGKSTSIQLLERFYDPDQGTVMIDGHDSKKVNIQFLRSNIGIVSQEPVLFDCSIMDNIKYGDNTKEISVERAIAAAKQAQLHDFVMSLPEKYETNVGIQGSQLSRGEKQRIAIARAIVRDPKILLLDEATSALDTESEKTVQTALDKAREGRTCIVIAHRLSTIQNSDIIAVVSQGVVIEKGTHEKLMAQKGAYYKLVITGAPIS q6psm0_rat Mdr1a protein protein rat Rattus norvegicus (Rat) Rat MELEEDLNGRADKNFSKMGKKSKKEKKEKKPAVSVLTMFRYAGWLDRFYMLLGTLAAIIHGIALPLMMLVFGDMTDSFANVGNNRSMSFYNATDIYAKLKDEMTTYAYYYTGIGAGVLIVAYIQVSLWCLAAGRQIHKIRQKFFHAIMNQEIGWFDVHDVGELNTRLTDDVSKINEGIGDKIGMFFQAMATFFGGFIIGFTRGWKLTLVILAISPVLGLSAGIWAKILSSFTDKELQAYAKAGAVAEEVLAAIRTVIAFGGQKKELERYNNNLEEAKRLGIKKAITANISMGAAFLLIYASYALAFWYGTSLVISKEYTIGQVLTVFFSVLIGAFSVGQASPNIEAFANARGAAYEVFSIIDNKPSIDSFSKSGHKPDNIQGNLEFKNIHFSYPSRKDVQILKGLNLKVKSGQTVALVGNSGCGKSTTVQLLQRLYDPIEGEVSIDGQDIRTINVRYLREIIGVVSQEPVLFATTIAENIRYGRENVTMDEIEKAVKEANAYDFIMKLPHKFDTLVGERGAQLSGGQKQRIAIARALVRNPKILLLDEATSALDTESEAVVQAALDKAREGRTTIVIAHRLSTVRNADVIAGFDGGVIVEQGNHDELMREKGIYFKLVMTQTAGNEIELGNEACESKDGIDNVDMSSKDSGSSLIRRRSTRKSIRGPHDQDGELSTKEALDDDVPPASFWRILKLNSTEWPYFVVGVFCAIINGGLQPAFSIIFSKVVGVFTKNDTPEIQRQNSNLFSLLFLILGIISFITFFLQGFTFGKAGEILTKRLRYMVFKSMLRQDISWFDDPKNTTGALTTRLANDAAQVKGATGSRLAVITQNIANLGTGIIISLIYGWQLTLLLLAIVPIIAIAGVVEMKMLSGQALKDKKELEGSGKIATEAIENFRTVVSLTREQKFETMYAQSLQIPYRNALKKAHVFGITFSFTQAMMYFSYAACFRFGAYLVARELMTFENVLLVFSAIVFGAMAVGQVSSFAPDYAKAKVSASHIIRIIEKIPEIDSYSTEGLKPNMLEGNVKFNGVMFNYPTRPNIPVLQGLSLEGKKGQTLALVGSSGCGKSTVVQLLERFYDPMAGTVFLDGKEIKQLNVQWLRAHLGIVSQEPILFDCSIAENIAYGDNSRVVSHEEIVKAAKEANIHQFIDSLPEKYNTRVGDKGTQLSGGQKQRIAIARALVRQPHILLLDEATSALDTESEKVVQEALDKAREGRTCIVIAHRLSTIQNADLIVVIQNGQVKEHGTHQQLLAQKGIYFSMVSVQAGAKRS hax1_human HCLS1-associated protein X-1 HAX1 HS1BP1 HS1-associating protein X-1 HS1-binding protein 1 protein protein human Homo sapiens Human MSLFDLFRGFFGFPGPRSHRDPFFGGMTRDEDDDEEEEEEGGSWGRGNPRFHSPQHPPEEFGFGFSFSPGGGIRFHDNFGFDDLVRDFNSIFSDMGAWTLPSHPPELPGPESETPGERLREGQTLRDSMLKYPDSHQPRIFGGVLESDARSESPQPAPDWGSQRPFHRFDDVWPMDPHPRTREDNDLDSQVSQEGLGPVLQPQPKSYFKSISVTKITKPDGIVEERRTVVDSEGRTETTVTRHEADSSPRGDPESPRPPALDDAFSILDLFLGRWFRSR o70420_rat protein protein rat Rattus norvegicus (Rat) Rat MWKASAGHAVSITQDDGEADDWETDPDFVNDVSEKEQRWGAKTVQGSGHQEHINIHKLRENVFQEHQTLKENELETGPKASHGYGGKFGVEQDRMDKSAVGHEYQSKLSKHCSQVDSVRGFGGKFGVQMDRVDQSAVGFEYQGKTEKHASQKDYSSGFGGKYGVQADRVDKSAVGFDYQGKTEKHESQKDYSKGFGGKYGIDKDKVDKSAVGFEYQGKTEKHESQKDYVKGFGGKFGVQTDRQDKCALGWDHQEKLQLHESQKDYAKGFGGKYGVQKDRMDKNASTFEEVVQVPSAYQKTVPIEAVTSKTSNIRANFESLAKEREQEDRRKAEAERAQRMAQERQEQEARRKLEEQARAKKQTPPASPSPQPAEDRPPSSPIYEDTAPLKAEPSYGSSEPEPEYSTEAAGLPEASNQQGLAYTSEPVYETTEVPGHYQAEDDTYDGYESDLGITAIALYDYQAAGDDEISFDPDDVITNIEMIDDGWWRGVCKGRYGLFPANYVELRQ mdr2-hax1-2 Y2H: bait = rat MDR2, prey = rat HAX1 365805 N/A 365811 unspecified role unspecified role bait bait linker domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false region required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain false N/A 365812 unspecified role unspecified role prey prey physical association physical association false false false bsep-hax1-1 Y2H: bait = rat BSEP, prey = rat HAX1 365805 N/A 365813 unspecified role unspecified role bait bait linker domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false N/A 365812 unspecified role unspecified role prey prey physical association physical association false false false mdr1a-hax1-1 Y2H: bait = rat Mdr1a, prey = rat HAX1 365805 N/A 365812 unspecified role unspecified role prey prey N/A 365814 unspecified role unspecified role bait bait linker domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false bsep-hax1-3 Co-IP : bait = rat BSEP, prey = rat HAX1 365806 N/A 365813 unspecified role unspecified role bait bait N/A 365812 unspecified role unspecified role prey prey physical association physical association false false false overexpressed. Q7TSE9. O70127. bsep-hax1-4 Co-IP : bait = rat BSEP, prey = human HAX1 365806 N/A 365815 unspecified role unspecified role prey prey N/A 365813 unspecified role unspecified role bait bait physical association physical association false false false overexpressed. O70127. mdr1a-hax1-2 Co-IP: bait = rat Mdr1a, prey = human HAX1 365806 N/A 365815 unspecified role unspecified role prey prey N/A 365814 unspecified role unspecified role bait bait 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 physical association physical association false false false over-expressed. Q6PSM0. bsep-hax1-7 Co-IP : bait = rat BSEP, prey = rat HAX1 365807 N/A 365812 unspecified role unspecified role prey prey N/A 365813 unspecified role unspecified role bait bait physical association physical association false false false mdr1a-hax1-3 Co-IP: bait = rat Mdr1a, prey = rat HAX1 365807 N/A 365814 unspecified role unspecified role bait bait N/A 365812 unspecified role unspecified role prey prey physical association physical association false false false The antibody used recognize both MDR1 and MDR2. Both MDR1a and MDR2 were shown to bind rat HAX-1 in vitro. cttn-hax1-8 Co-IP : bait = rat cortactin prey = rat HAX1 365807 N/A 365816 unspecified role unspecified role bait bait N/A 365812 unspecified role unspecified role prey prey physical association physical association false false false The antibody my not be specific to an isofom of cortactin. mdr2-hax1-3 GST pull-down: bait = rat MDR2, prey = rat HAX1 365808 N/A 365812 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver N/A 365811 unspecified role unspecified role bait bait linker domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 bsep-hax1-2 GST pull-down : bait = rat BSEP, prey = rat HAX1 365808 N/A 365813 unspecified role unspecified role bait bait linker domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 365812 unspecified role unspecified role prey prey rat-liver rattus norvegicus liver Rat liver liver physical association physical association false false false mdr2-hax1-1 Y2H: bait = rat MDR2, prey = rat HAX1 365809 N/A 365812 unspecified role unspecified role prey prey N/A 365811 unspecified role unspecified role bait bait linker domain sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false bsep-hax1-6 Rat BSEP and rat HAX1 colocalize in the apical domain of MDCK cells 365810 N/A 365813 unspecified role unspecified role neutral component neutral component eyfp yfp tag yellow fluorescent protein tag YFP yellow fluorescent protein yellow fluorescent protein YFP yfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 365812 unspecified role unspecified role neutral component neutral component colocalization colocalization false false false overexpressed. Q7TSE9. O70127. One to 2 days after confluency, cells showed differentiation of apical and lateral membranes. HAX-1 partially co-localized with BSEP-EYFP in the apical membrane; however, most HAX-1 was located immediately below the apical domain, where it partially localized with BSEP-EYFP. In polarized columnar cells, 4-5 days after confluency, BSEP-EYFP was mostly restricted to the apical membrane where it co-localized with HAX-1