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/ jeong-2009-3 Human selenium binding protein-1 (hSP56) interacts with VDU1 in a selenium-dependent manner. in vitro In vitro elisa enzyme linked immunosorbent assay ELISA ELISA elisa antibody detection identification by antibody antibody detection Jeong JY., Wang Y., Sytkowski AJ. asytkows@bidmc.harvard.edu Human prostate cDNA library. Biochem. Biophys. Res. Commun. (0006-291X) 2009 Only protein-protein interactions imex curation jeong-2009-1 Human selenium binding protein-1 (hSP56) interacts with VDU1 in a selenium-dependent manner. 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 Jeong JY., Wang Y., Sytkowski AJ. asytkows@bidmc.harvard.edu Human prostate cDNA library. Biochem. Biophys. Res. Commun. (0006-291X) 2009 Only protein-protein interactions imex curation jeong-2009-2 Human selenium binding protein-1 (hSP56) interacts with VDU1 in a selenium-dependent manner. in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Jeong JY., Wang Y., Sytkowski AJ. asytkows@bidmc.harvard.edu Human prostate cDNA library. Biochem. Biophys. Res. Commun. (0006-291X) 2009 Only protein-protein interactions imex curation jeong-2009-4 Human selenium binding protein-1 (hSP56) interacts with VDU1 in a selenium-dependent manner. human-lncap Human caucasian prostate carcinoma lncap Human caucasian prostate adenocarcinoma fluorescence imaging fluorescence microscopy fluorescence imaging anti tag immunost anti tag immunostaining anti tag immunost Jeong JY., Wang Y., Sytkowski AJ. asytkows@bidmc.harvard.edu Human prostate cDNA library. Biochem. Biophys. Res. Commun. (0006-291X) 2009 Only protein-protein interactions imex curation jeong-2009-5 Human selenium binding protein-1 (hSP56) interacts with VDU1 in a selenium-dependent manner. in vitro In vitro comigration in sds comigration in sds page comigration in sds weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Jeong JY., Wang Y., Sytkowski AJ. asytkows@bidmc.harvard.edu Human prostate cDNA library. Biochem. Biophys. Res. Commun. (0006-291X) 2009 Only protein-protein interactions imex curation jeong-2009-6 Human selenium binding protein-1 (hSP56) interacts with VDU1 in a selenium-dependent manner. 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 Jeong JY., Wang Y., Sytkowski AJ. asytkows@bidmc.harvard.edu Human prostate cDNA library. Biochem. Biophys. Res. Commun. (0006-291X) 2009 Only protein-protein interactions imex curation q8tey7-2 Ubiquitin carboxyl-terminal hydrolase 33 USP33 KIAA1097 VDU1 Deubiquitinating enzyme 33 Ubiquitin-specific-processing protease 33 VHL-interacting deubiquitinating enzyme 1 Ubiquitin thioesterase 33 protein protein human Homo sapiens Human MSAFRNHCPHLDSVGEITKEDLIQKSLGTCQDCKVQGPNLWACLENRCSYVGCGESQVDHSTIHSQETKHYLTVNLTTLRVWCYACSKEVFLDRKLGTQPSLPHVRQPHQIQENSVQDFKIPSNTTLKTPLVAVFDDLDIEADEEDELRARGLTGLKNIGNTCYMNAALQALSNCPPLTQFFLDCGGLARTDKKPAICKSYLKLMTELWHKSRPGSVVPTTLFQGIKTVNPTFRGYSQQDAQEFLRCLMDLLHEELKEQVMEVEEDPQTITTEETMEEDKSQSDVDFQSCESCSNSDRAENENGSRCFSEDNNETTMLIQDDENNSEMSKDWQKEKMCNKINKVNSEGEFDKDRDSISETVDLNNQETVKVQIHSRASEYITDVHSNDLSTPQILPSNEGVNPRLSASPPKSGNLWPGLAPPHKKAQSASPKRKKQHKKYRSVISDIFDGTIISSVQCLTCDRVSVTLETFQDLSLPIPGKEDLAKLHSSSHPTSIVKAGSCGEAYAPQGWIAFFMEYVKRFVVSCVPSWFWGPVVTLQDCLAAFFARDELKGDNMYSCEKCKKLRNGVKFCKVQNFPEILCIHLKRFRHELMFSTKISTHVSFPLEGLDLQPFLAKDSPAQIVTYDLLSVICHHGTASSGHYIAYCRNNLNNLWYEFDDQSVTEVSESTVQNAEAYVLFYRKSSEEAQKERRRISNLLNIMEPSLLQFYISRQWLNKFKTFAEPGPISNNDFLCIHGGVPPRKAGYIEDLVLMLPQNIWDNLYSRYGGGPAVNHLYICHTCQIEAEKIEKRRKTELEIFIRLNRAFQKEDSPATFYCISMQWFREWESFVKGKDGDPPGPIDNTKIAVTKCGNVMLRQGADSGQISEETWNFLQSIYGGGPEVILRPPVVHVDPDILQAEEKIEVETRSL sbp1_human Methanethiol oxidase SELENBP1 SBP 56 kDa selenium-binding protein Selenium-binding protein 1 protein protein human Homo sapiens Human MATKCGNCGPGYSTPLEAMKGPREEIVYLPCIYRNTGTEAPDYLATVDVDPKSPQYCQVIHRLPMPNLKDELHHSGWNTCSSCFGDSTKSRTKLVLPSLISSRIYVVDVGSEPRAPKLHKVIEPKDIHAKCELAFLHTSHCLASGEVMISSLGDVKGNGKGGFVLLDGETFEVKGTWERPGGAAPLGYDFWYQPRHNVMISTEWAAPNVLRDGFNPADVEAGLYGSHLYVWDWQRHEIVQTLSLKDGLIPLEIRFLHNPDAAQGFVGCALSSTIQRFYKNEGGTWSVEKVIQVPPKKVKGWLLPEMPGLITDILLSLDDRFLYFSNWLHGDLRQYDISDPQRPRLTGQLFLGGSIVKGGPVQVLEDEELKSQPEPLVVKGKRVAGGPQMIQLSLDGKRLYITTSLYSAWDKQFYPDLIREGSVMLQVDVDTVKGGLKLNPNFLVDFGKEPLGPALAHELRYPGGDCSSDIWI ubp33_human Ubiquitin carboxyl-terminal hydrolase 33 Ubiquitin-specific-processing protease 33 Deubiquitinating enzyme 33 VHL-interacting deubiquitinating enzyme 1 USP33 KIAA1097 VDU1 Ubiquitin thioesterase 33 protein protein human Homo sapiens Human MTGSNSHITILTLKVLPHFESLGKQEKIPNKMSAFRNHCPHLDSVGEITKEDLIQKSLGTCQDCKVQGPNLWACLENRCSYVGCGESQVDHSTIHSQETKHYLTVNLTTLRVWCYACSKEVFLDRKLGTQPSLPHVRQPHQIQENSVQDFKIPSNTTLKTPLVAVFDDLDIEADEEDELRARGLTGLKNIGNTCYMNAALQALSNCPPLTQFFLDCGGLARTDKKPAICKSYLKLMTELWHKSRPGSVVPTTLFQGIKTVNPTFRGYSQQDAQEFLRCLMDLLHEELKEQVMEVEEDPQTITTEETMEEDKSQSDVDFQSCESCSNSDRAENENGSRCFSEDNNETTMLIQDDENNSEMSKDWQKEKMCNKINKVNSEGEFDKDRDSISETVDLNNQETVKVQIHSRASEYITDVHSNDLSTPQILPSNEGVNPRLSASPPKSGNLWPGLAPPHKKAQSASPKRKKQHKKYRSVISDIFDGTIISSVQCLTCDRVSVTLETFQDLSLPIPGKEDLAKLHSSSHPTSIVKAGSCGEAYAPQGWIAFFMEYVKRFVVSCVPSWFWGPVVTLQDCLAAFFARDELKGDNMYSCEKCKKLRNGVKFCKVQNFPEILCIHLKRFRHELMFSTKISTHVSFPLEGLDLQPFLAKDSPAQIVTYDLLSVICHHGTASSGHYIAYCRNNLNNLWYEFDDQSVTEVSESTVQNAEAYVLFYRKSSEEAQKERRRISNLLNIMEPSLLQFYISRQWLNKFKTFAEPGPISNNDFLCIHGGVPPRKAGYIEDLVLMLPQNIWDNLYSRYGGGPAVNHLYICHTCQIEAEKIEKRRKTELEIFIRLNRAFQKEDSPATFYCISMQWFREWESFVKGKDGDPPGPIDNTKIAVTKCGNVMLRQGADSGQISEETWNFLQSIYGGGPEVILRPPVVHVDPDILQAEEKIEVETRSL se Selenium small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) hsbp1-vdu1-3 867822 n/a 867828 unspecified role unspecified role prey prey c-terminal region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n-terminal 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 867829 unspecified role unspecified role bait bait direct interaction direct interaction false false false Fig 1C. hsbp1-vdu1-se 867822 n/a 867828 unspecified role unspecified role prey prey c-terminal ha-tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false n/a 867829 unspecified role unspecified role bait bait chlae-cos_7 Cercopithecus aethiops simian cells transformed with SV40 Grivet cos_7 cercopithecus aethiops, by transformation with SV40 physical association physical association false false false This interaction only occurred if hSBP1 was pre-incubated with sodium selenite for 30 mins at RT. Similarly, a reaction only took place under non-reducing conditions i.e. the absence of 2-ME. Fig 2C. Selenium selenite hsbp1-vdu1 867823 n/a 867830 unspecified role unspecified role prey prey c-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n/a 867829 unspecified role unspecified role bait bait physical association physical association false false false hsbp1-vdu1-2 867824 n/a 867828 unspecified role unspecified role bait bait c-terminal region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false n-terminal 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 867829 unspecified role unspecified role prey prey direct interaction direct interaction false false false Fig 1B. hsbp1-vdu1-1 867825 n/a 867828 unspecified role unspecified role neutral component neutral component c-terminal ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false n/a 867829 unspecified role unspecified role neutral component neutral component c-terminal egfp tag enhanced green fluorescent protein tag enhanced green fluorescent protein EGFP enhanced green fluorescent protein EGFP egfp tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range false colocalization colocalization false false false Fig 1D. selenbp1-se 867826 n/a 867831 unspecified role unspecified role neutral component neutral component n/a 867829 unspecified role unspecified role neutral component neutral component direct interaction direct interaction false false false Selenium binding to hSP56 was only maintained under non-reducing conditions. Radiolabelled sodium selenite (75-Selenium) was used in SDS-PAGE. sbp1-vdu1 867827 n/a 867829 unspecified role unspecified role bait bait n/a 867828 unspecified role unspecified role prey prey c-terminal region 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 Fig. 1A We isolated the library plasmid encoding the VDU1 C-terminal fragment (AD-VDU1f) and examined its interaction with hSP56 using yeast two-hybrid analysis (Fig. 1A)