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/ V-ATPase interacts with ARNO and Arf6 in early endosomes and regulates the protein degradative pathway. V-ATPase interacts with ARNO and Arf6 in early endosomes and regulates the protein degradative pathway. Nat. Cell Biol. (1465-7392) 2006 imex curation Hurtado-Lorenzo A., Skinner M., El Annan J., Futai M., Sun-Wada GH., Bourgoin S., Casanova J., Wildeman A., Bechoua S., Ausiello DA., Brown D., Marshansky V. Synapse - Interactions of proteins with an established role in the presynapse. BioCreative - Critical Assessment of Information Extraction systems in Biology Vladmir_Marshansky@hms.harvard.edu 24-JAN-2007: Contacted by JYOTI. Only protein-protein interactions 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. 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 Hurtado-Lorenzo A., Skinner M., El Annan J., Futai M., Sun-Wada GH., Bourgoin S., Casanova J., Wildeman A., Bechoua S., Ausiello DA., Brown D., Marshansky V. Synapse - Interactions of proteins with an established role in the presynapse. Vladmir_Marshansky@hms.harvard.edu Accepted 2006-SEP-1 by DAVET. 24-JAN-2007: Contacted by JYOTI. Nat. Cell Biol. (1465-7392) 2006 Only protein-protein interactions imex curation V-ATPase interacts with ARNO and Arf6 in early endosomes and regulates the protein degradative pathway. V-ATPase interacts with ARNO and Arf6 in early endosomes and regulates the protein degradative pathway. Nat. Cell Biol. (1465-7392) 2006 imex curation Hurtado-Lorenzo A., Skinner M., El Annan J., Futai M., Sun-Wada GH., Bourgoin S., Casanova J., Wildeman A., Bechoua S., Ausiello DA., Brown D., Marshansky V. Synapse - Interactions of proteins with an established role in the presynapse. BioCreative - Critical Assessment of Information Extraction systems in Biology Vladmir_Marshansky@hms.harvard.edu 24-JAN-2007: Contacted by JYOTI. Only protein-protein interactions human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Hurtado-Lorenzo A., Skinner M., El Annan J., Futai M., Sun-Wada GH., Bourgoin S., Casanova J., Wildeman A., Bechoua S., Ausiello DA., Brown D., Marshansky V. Synapse - Interactions of proteins with an established role in the presynapse. Vladmir_Marshansky@hms.harvard.edu Accepted 2006-SEP-1 by DAVET. 24-JAN-2007: Contacted by JYOTI. Nat. Cell Biol. (1465-7392) 2006 Only protein-protein interactions imex curation V-ATPase interacts with ARNO and Arf6 in early endosomes and regulates the protein degradative pathway. V-ATPase interacts with ARNO and Arf6 in early endosomes and regulates the protein degradative pathway. Nat. Cell Biol. (1465-7392) 2006 imex curation Hurtado-Lorenzo A., Skinner M., El Annan J., Futai M., Sun-Wada GH., Bourgoin S., Casanova J., Wildeman A., Bechoua S., Ausiello DA., Brown D., Marshansky V. Synapse - Interactions of proteins with an established role in the presynapse. BioCreative - Critical Assessment of Information Extraction systems in Biology Vladmir_Marshansky@hms.harvard.edu 24-JAN-2007: Contacted by JYOTI. Only protein-protein interactions 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 tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Hurtado-Lorenzo A., Skinner M., El Annan J., Futai M., Sun-Wada GH., Bourgoin S., Casanova J., Wildeman A., Bechoua S., Ausiello DA., Brown D., Marshansky V. Synapse - Interactions of proteins with an established role in the presynapse. Vladmir_Marshansky@hms.harvard.edu Accepted 2006-SEP-1 by DAVET. 24-JAN-2007: Contacted by JYOTI. Nat. Cell Biol. (1465-7392) 2006 Only protein-protein interactions imex curation V-ATPase interacts with ARNO and Arf6 in early endosomes and regulates the protein degradative pathway. V-ATPase interacts with ARNO and Arf6 in early endosomes and regulates the protein degradative pathway. Nat. Cell Biol. (1465-7392) 2006 imex curation Hurtado-Lorenzo A., Skinner M., El Annan J., Futai M., Sun-Wada GH., Bourgoin S., Casanova J., Wildeman A., Bechoua S., Ausiello DA., Brown D., Marshansky V. Synapse - Interactions of proteins with an established role in the presynapse. BioCreative - Critical Assessment of Information Extraction systems in Biology Vladmir_Marshansky@hms.harvard.edu 24-JAN-2007: Contacted by JYOTI. Only protein-protein interactions 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 Hurtado-Lorenzo A., Skinner M., El Annan J., Futai M., Sun-Wada GH., Bourgoin S., Casanova J., Wildeman A., Bechoua S., Ausiello DA., Brown D., Marshansky V. Synapse - Interactions of proteins with an established role in the presynapse. Vladmir_Marshansky@hms.harvard.edu Accepted 2006-SEP-1 by DAVET. 24-JAN-2007: Contacted by JYOTI. Nat. Cell Biol. (1465-7392) 2006 Only protein-protein interactions imex curation V-ATPase interacts with ARNO and Arf6 in early endosomes and regulates the protein degradative pathway. V-ATPase interacts with ARNO and Arf6 in early endosomes and regulates the protein degradative pathway. Nat. Cell Biol. (1465-7392) 2006 imex curation Hurtado-Lorenzo A., Skinner M., El Annan J., Futai M., Sun-Wada GH., Bourgoin S., Casanova J., Wildeman A., Bechoua S., Ausiello DA., Brown D., Marshansky V. Synapse - Interactions of proteins with an established role in the presynapse. BioCreative - Critical Assessment of Information Extraction systems in Biology Vladmir_Marshansky@hms.harvard.edu 24-JAN-2007: Contacted by JYOTI. Only protein-protein interactions human-mtc Human medullary thyroid carcinoma mtc Human medullary thyroid carcinoma anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Hurtado-Lorenzo A., Skinner M., El Annan J., Futai M., Sun-Wada GH., Bourgoin S., Casanova J., Wildeman A., Bechoua S., Ausiello DA., Brown D., Marshansky V. Synapse - Interactions of proteins with an established role in the presynapse. Vladmir_Marshansky@hms.harvard.edu Accepted 2006-SEP-1 by DAVET. 24-JAN-2007: Contacted by JYOTI. Nat. Cell Biol. (1465-7392) 2006 Only protein-protein interactions imex curation V-ATPase interacts with ARNO and Arf6 in early endosomes and regulates the protein degradative pathway. V-ATPase interacts with ARNO and Arf6 in early endosomes and regulates the protein degradative pathway. Nat. Cell Biol. (1465-7392) 2006 imex curation Hurtado-Lorenzo A., Skinner M., El Annan J., Futai M., Sun-Wada GH., Bourgoin S., Casanova J., Wildeman A., Bechoua S., Ausiello DA., Brown D., Marshansky V. Synapse - Interactions of proteins with an established role in the presynapse. BioCreative - Critical Assessment of Information Extraction systems in Biology Vladmir_Marshansky@hms.harvard.edu 24-JAN-2007: Contacted by JYOTI. Only protein-protein interactions mouse-prox_tubule_cell Mouse proximal tubule cells kidney-proximal_tubule Proximal Tubule cell anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Hurtado-Lorenzo A., Skinner M., El Annan J., Futai M., Sun-Wada GH., Bourgoin S., Casanova J., Wildeman A., Bechoua S., Ausiello DA., Brown D., Marshansky V. Synapse - Interactions of proteins with an established role in the presynapse. Vladmir_Marshansky@hms.harvard.edu Accepted 2006-SEP-1 by DAVET. Nat. Cell Biol. (1465-7392) 2006 Only protein-protein interactions imex curation V-ATPase interacts with ARNO and Arf6 in early endosomes and regulates the protein degradative pathway. V-ATPase interacts with ARNO and Arf6 in early endosomes and regulates the protein degradative pathway. Nat. Cell Biol. (1465-7392) 2006 imex curation Hurtado-Lorenzo A., Skinner M., El Annan J., Futai M., Sun-Wada GH., Bourgoin S., Casanova J., Wildeman A., Bechoua S., Ausiello DA., Brown D., Marshansky V. Synapse - Interactions of proteins with an established role in the presynapse. BioCreative - Critical Assessment of Information Extraction systems in Biology Vladmir_Marshansky@hms.harvard.edu 24-JAN-2007: Contacted by JYOTI. Only protein-protein interactions in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Hurtado-Lorenzo A., Skinner M., El Annan J., Futai M., Sun-Wada GH., Bourgoin S., Casanova J., Wildeman A., Bechoua S., Ausiello DA., Brown D., Marshansky V. Synapse - Interactions of proteins with an established role in the presynapse. Vladmir_Marshansky@hms.harvard.edu Accepted 2006-SEP-1 by DAVET. 24-JAN-2007: Contacted by JYOTI. Nat. Cell Biol. (1465-7392) 2006 Only protein-protein interactions imex curation V-ATPase interacts with ARNO and Arf6 in early endosomes and regulates the protein degradative pathway. V-ATPase interacts with ARNO and Arf6 in early endosomes and regulates the protein degradative pathway. Nat. Cell Biol. (1465-7392) 2006 imex curation Hurtado-Lorenzo A., Skinner M., El Annan J., Futai M., Sun-Wada GH., Bourgoin S., Casanova J., Wildeman A., Bechoua S., Ausiello DA., Brown D., Marshansky V. Synapse - Interactions of proteins with an established role in the presynapse. BioCreative - Critical Assessment of Information Extraction systems in Biology Vladmir_Marshansky@hms.harvard.edu 24-JAN-2007: Contacted by JYOTI. Only protein-protein interactions human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Hurtado-Lorenzo A., Skinner M., El Annan J., Futai M., Sun-Wada GH., Bourgoin S., Casanova J., Wildeman A., Bechoua S., Ausiello DA., Brown D., Marshansky V. Synapse - Interactions of proteins with an established role in the presynapse. Vladmir_Marshansky@hms.harvard.edu Accepted 2006-SEP-1 by DAVET. 24-JAN-2007: Contacted by JYOTI. Nat. Cell Biol. (1465-7392) 2006 Only protein-protein interactions imex curation cyh2_human Cytohesin-2 PH, SEC7 and coiled-coil domain-containing protein 2 ARF nucleotide-binding site opener ARF exchange factor CYTH2 ARNO PSCD2L PSCD2 protein protein human Homo sapiens Human MEDGVYEPPDLTPEERMELENIRRRKQELLVEIQRLREELSEAMSEVEGLEANEGSKTLQRNRKMAMGRKKFNMDPKKGIQFLVENELLQNTPEEIARFLYKGEGLNKTAIGDYLGEREELNLAVLHAFVDLHEFTDLNLVQALRQFLWSFRLPGEAQKIDRMMEAFAQRYCLCNPGVFQSTDTCYVLSFAVIMLNTSLHNPNVRDKPGLERFVAMNRGINEGGDLPEELLRNLYDSIRNEPFKIPEDDGNDLTHTFFNPDREGWLLKLGGGRVKTWKRRWFILTDNCLYYFEYTTDKEPRGIIPLENLSIREVDDPRKPNCFELYIPNNKGQLIKACKTEADGRVVEGNHMVYRISAPTQEEKDEWIKSIQAAVSVDPFYEMLAARKKRISVKKKQEQP DIP protein Q99418 original sequence version: 131 DIP protein Q99418 original sequence version: 140 DIP protein Q99418-1 original sequence version: 125 DIP protein Q99418 original sequence version: 125 70441A58483BD0E1 arf6_mouse ADP-ribosylation factor 6 Arf6 protein protein mouse Mus musculus Mouse MGKVLSKIFGNKEMRILMLGLDAAGKTTILYKLKLGQSVTTIPTVGFNVETVTYKNVKFNVWDVGGQDKIRPLWRHYYTGTQGLIFVVDCADRDRIDEARQELHRIINDREMRDAIILIFANKQDLPDAMKPHEIQEKLGLTRIRDRNWYVQPSCATSGDGLYEGLTWLTSNYKS 49E38E59AEA52B98 vatc1_human V-type proton ATPase subunit C 1 Vacuolar proton pump subunit C 1 ATP6V1C1 ATP6C ATP6D VATC protein protein human Homo sapiens Human MTEFWLISAPGEKTCQQTWEKLHAATSKNNNLAVTSKFNIPDLKVGTLDVLVGLSDELAKLDAFVEGVVKKVAQYMADVLEDSKDKVQENLLANGVDLVTYITRFQWDMAKYPIKQSLKNISEIIAKGVTQIDNDLKSRASAYNNLKGNLQNLERKNAGSLLTRSLAEIVKKDDFVLDSEYLVTLLVVVPKLNHNDWIKQYETLAEMVVPRSSNVLSEDQDSYLCNVTLFRKAVDDFRHKARENKFIVRDFQYNEEEMKADKEEMNRLSTDKKKQFGPLVRWLKVNFSEAFIAWIHVKALRVFVESVLRYGLPVNFQAMLLQPNKKTLKKLREVLHELYKHLDSSAAAIIDAPMDIPGLNLSQQEYYPYVYYKIDCNLLEFK 5626E2AB2BD66BA7 arf6_human ADP-ribosylation factor 6 ARF6 protein protein human Homo sapiens Human MGKVLSKIFGNKEMRILMLGLDAAGKTTILYKLKLGQSVTTIPTVGFNVETVTYKNVKFNVWDVGGQDKIRPLWRHYYTGTQGLIFVVDCADRDRIDEARQELHRIINDREMRDAIILIFANKQDLPDAMKPHEIQEKLGLTRIRDRNWYVQPSCATSGDGLYEGLTWLTSNYKS DIP protein P62330 original sequence version: 126 DIP protein P62330 original sequence version: 113 DIP protein P62330 original sequence version: 105 49E38E59AEA52B98 vpp2_human V-type proton ATPase 116 kDa subunit a2 Vacuolar proton translocating ATPase 116 kDa subunit a isoform 2 Lysosomal H(+)-transporting ATPase V0 subunit a2 TJ6 ATP6V0A2 protein protein human Homo sapiens Human MGSLFRSETMCLAQLFLQSGTAYECLSALGEKGLVQFRDLNQNVSSFQRKFVGEVKRCEELERILVYLVQEINRADIPLPEGEASPPAPPLKQVLEMQEQLQKLEVELREVTKNKEKLRKNLLELIEYTHMLRVTKTFVKRNVEFEPTYEEFPSLESDSLLDYSCMQRLGAKLGFVSGLINQGKVEAFEKMLWRVCKGYTIVSYAELDESLEDPETGEVIKWYVFLISFWGEQIGHKVKKICDCYHCHVYPYPNTAEERREIQEGLNTRIQDLYTVLHKTEDYLRQVLCKAAESVYSRVIQVKKMKAIYHMLNMCSFDVTNKCLIAEVWCPEADLQDLRRALEEGSRESGATIPSFMNIIPTKETPPTRIRTNKFTEGFQNIVDAYGVGSYREVNPALFTIITFPFLFAVMFGDFGHGFVMFLFALLLVLNENHPRLNQSQEIMRMFFNGRYILLLMGLFSVYTGLIYNDCFSKSVNLFGSGWNVSAMYSSSHPPAEHKKMVLWNDSVVRHNSILQLDPSIPGVFRGPYPLGIDPIWNLATNRLTFLNSFKMKMSVILGIIHMTFGVILGIFNHLHFRKKFNIYLVSIPELLFMLCIFGYLIFMIFYKWLVFSAETSRVAPSILIEFINMFLFPASKTSGLYTGQEYVQRVLLVVTALSVPVLFLGKPLFLLWLHNGRSCFGVNRSGYTLIRKDSEEEVSLLGSQDIEEGNHQVEDGCREMACEEFNFGEILMTQVIHSIEYCLGCISNTASYLRLWALSLAHAQLSDVLWAMLMRVGLRVDTTYGVLLLLPVIALFAVLTIFILLIMEGLSAFLHAIRLHWVEFQNKFYVGAGTKFVPFSFSLLSSKFNNDDSVA 6C38B36FA33A92BA vatc1_mouse V-type proton ATPase subunit C 1 Vacuolar proton pump subunit C 1 Atp6v1c1 Atp6c Vatc Atp6c1 protein protein mouse Mus musculus Mouse MTEFWLISAPGEKTCQQTWEKLHAATTKNNNLAVSSKFNIPDLKVGTLDVLVGLSDELAKLDAFVEGVVKKVAQYMADVLEDSKDKVQENLLASGVDLVTYITRFQWDMAKYPIKQSLKNISEIIAKGVTQIDNDLKSRASAYNNLKGNLQNLERKNAGSLLTRSLAEIVKKDDFVLDSEYLVTLLVVVPKLNHNDWIKQYETLAEMVVPRSSNVLSEDQDSYLCNVTLFRKAVDDFRHKARENKFIVRDFQYNEEEMKADKEEMTRLSTDKKKQFGPLVRWLKVNFSEAFIAWIHIKALRVFVESVLRYGLPVNFQAMLLQPNKKSVKKLREVLHELYKHLDSSAAAIIDAPMDIPGLNLSQQEYYPYVYYKIDCNLLEFK 6D1851D8C81DF98F vpp2_mouse V-type proton ATPase 116 kDa subunit a2 Vacuolar proton translocating ATPase 116 kDa subunit a isoform 2 Lysosomal H(+)-transporting ATPase V0 subunit a2 Immune suppressor factor J6B7 ShIF Atp6v0a2 Atp6n1b Tj6 protein protein mouse Mus musculus Mouse MGSLFRSESMCLAQLFLQSGTAYECLSALGEKGLVQFRDLNQNVSSFQRKFVGEVKRCEELERILVYLVQEITRADIPLPEGEASPPAPPLKHVLEMQEQLQKLEVELREVTKNKEKLRKNLLELVEYTHMLRVTKTFLKRNVEFEPTYEEFPALENDSLLDYSCMQRLGAKLGFVSGLIQQGRVEAFERMLWRACKGYTIVTYAELDECLEDPETGEVIKWYVFLISFWGEQIGHKVKKICDCYHCHIYPYPNTAEERREIQEGLNTRIQDLYTVLHKTEDYLRQVLCKAAESVCSRVVQVRKMKAIYHMLNMCSFDVTNKCLIAEVWCPEVDLPGLRRALEEGSRESGATIPSFMNTIPTKETPPTLIRTNKFTEGFQNIVDAYGVGSYREVNPALFTIITFPFLFAVMFGDFGHGFVMFLFALLLVLNENHPRLSQSQEILRMFFDGRYILLLMGLFSVYTGLIYNDCFSKSVNLFGSGWNVSAMYSSSHSPEEQRKMVLWNDSTIRHSRTLQLDPNIPGVFRGPYPFGIDPIWNLATNRLTFLNSFKMKMSVILGIFHMTFGVVLGIFNHLHFRKKFNVYLVSVPEILFMLCIFGYLIFMIIYKWLAYSAETSREAPSILIEFINMFLFPTSKTHGLYPGQAHVQRVLVALTVLAVPVLFLGKPLFLLWLHNGRNCFGMSRSGYTLVRKDSEEEVSLLGNQDIEEGNSRMEEGCREVTCEEFNFGEILMTQAIHSIEYCLGCISNTASYLRLWALSLAHAQLSDVLWAMLMRVGLRVDTTYGVLLLLPVMAFFAVLTIFILLVMEGLSAFLHAIRLHWVEFQNKFYVGAGTKFVPFSFSLLSSKFSNDDSIA 6A0D593F6F401E22 cyh2_mouse Cytohesin-2 PH, SEC7 and coiled-coil domain-containing protein 2 SEC7 homolog B Pscd2 Cyth2 Sec7b ARF nucleotide-binding site opener protein protein mouse Mus musculus Mouse MEDGVYEPPDLTPEERMELENIRRRKQELLVEIQRLREELSEAMSEVEGLEANEGSKTLQRNRKMAMGRKKFNMDPKKGIQFLVEHELLQNTPEEIARFLYKGEGLNKTAIGDYLGEREELNLSVLHAFVDLHEFTDLNLVQALRQFLWSFRLPGEAQKIDRMMEAFAQRYCLCNPGVFQSTDTCYVLSFAVIMLNTSLHNPNVRDKPGLERFVAMNRGINEGGDLPEDLLRNLYDSIRNEPFKIPEDDGNDLTHTFFNPDREGWLLKLAGGRVKTWKRRWFILTDNCLYYFEYTTDKEPRGIIPLENLSIREVDDPRKPNCFELYIPNNKGQLIKACKTEADGRVVEGNHMVYRISAPTQEEKDEWIKSIQAAVSVDPFYEMLAARKKRISVKKKQEQP 8F6F4F7C2F931A89 pscd2-atp6v0a2-1 6 15 unspecified role unspecified role bait bait 16 unspecified role unspecified role prey prey physical association physical association 3A, 3G, 4F Western blot (WB) showing immunoprecipitation (IP) of ARNO with the a2-isoform from proximal tubule post-mitochondrial supernatant (PMS) Process is acidification dependent FCCP - an uncoupler of endosomal acidification pscd2-atp6v0a2-2 7 15 unspecified role unspecified role prey prey mouse-prox_tubule_cell Mouse proximal tubule cells kidney-proximal_tubule Proximal Tubule cell 16 unspecified role unspecified role bait bait 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 ecoli Escherichia coli (strain K12) physical association physical association 3B Pull-down (PD) of the a2-isoform with recombinant ARNO(WT)-6His used as bait from (b) proximal tubule PMS and (c) isolated endosomes (Endo). pscd2-atp6v0a2-3 7 16 unspecified role unspecified role bait bait 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 ecoli Escherichia coli (strain K12) 15 unspecified role unspecified role prey prey mouse-endosome Mouse endosome endosome Endosome physical association physical association 3C Pull-down (PD) of the a2-isoform with recombinant ARNO(WT)-6His used as bait from (b) proximal tubule PMS and (c) isolated endosomes (Endo). M&M implies endosomes from rat or dog, text implies mouse. I have gone with mouse since that is organism used throughout for everything else. pscd2-atp6v0a2-4 7 15 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro 16 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 region sufficient to bind sufficient binding region sufficient to bind range range certain certain sequence position certain ecoli Escherichia coli (strain K12) direct interaction direct interaction 3D & 3E Autoradiography (AR) showing the direct interacion of a2N-35S with GST-ARNO(WT) d) via carboxy-terminus but not amino terminus of ARNO (e) pscd2-atp6v0a2-5 1 16 unspecified role unspecified role prey prey ecoli Escherichia coli (strain K12) 15 unspecified role unspecified role bait bait mouse-endosome Mouse endosome endosome Endosome physical association physical association 3F Mammalian two-hybrid (M2H) assay demonstrating direct interaction of a2N(pMa2N) with ARNO(pVARNO) in HEK cells. pscd2-atp6v0a2-6 2 13 unspecified role unspecified role bait bait 9 unspecified role unspecified role prey prey physical association physical association 3G Western blot showing immunoprecipitation of ARNO (upper panel) but not of Arf6 (lower panel) with the a2-isoform of vesicles isolated from MTC, HeLa and HEK cells. pscd2-atp6v0a2-7 4 13 unspecified role unspecified role bait bait 9 unspecified role unspecified role prey prey physical association physical association 3G Western blot showing immunoprecipitation of ARNO (upper panel) but not of Arf6 (lower panel) with the a2-isoform of vesicles isolated from MTC, HeLa and HEK cells. arf6-atp6v1c1-1 3 10 unspecified role unspecified role prey prey region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 11 unspecified role unspecified role bait bait hsv tagged region tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association 3H Western blot analysis (right panel) showing co-immunoprecipitation and interaction of Arf6-HA with c-HSV in HEK cells. arf6-atp6v1c1-2 2 11 unspecified role unspecified role bait bait 12 unspecified role unspecified role prey prey physical association physical association 3I & 3J Western blot analysis showing co-immunoprecipitation and interaction of endogenous Arf6 with the c-subunit of V-ATPase. arf6-atp6v1c1-3 6 14 unspecified role unspecified role bait bait 10 unspecified role unspecified role prey prey physical association physical association 3I & 3J Western blot analysis showing co-immunoprecipitation and interaction of endogenous Arf6 with the c-subunit of V-ATPase. arf6-atp6v1c1-4 4 12 unspecified role unspecified role prey prey 11 unspecified role unspecified role bait bait physical association physical association 3I & 3J Western blot analysis showing co-immunoprecipitation and interaction of endogenous Arf6 with the c-subunit of V-ATPase. arf6-atp6v1c1-5 5 12 unspecified role unspecified role prey prey 11 unspecified role unspecified role bait bait hsv tagged region tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association 3I, 3J Western blot analysis using anti-Arf6 antibody shows the specific co-immunoprecipitation of endogenous Arf6 (upper panel) but not Arf1 (lower panel) with the c-subunit from MTC, HeLa and HEK vesicular fractions. (i, j) arf6-atp6v1c1-6 8 12 unspecified role unspecified role prey prey 11 unspecified role unspecified role bait bait hsv tagged region tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association 3I, 3J Western blot analysis using anti-Arf6 antibody shows the specific co-immunoprecipitation of endogenous Arf6 (upper panel) but not Arf1 (lower panel) with the c-subunit from MTC, HeLa and HEK vesicular fractions. (i, j)