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/ Arabidopsis Protein Disulfide Isomerase-5 Inhibits Cysteine Proteases during Trafficking to Vacuoles before Programmed Cell Death of the Endothelium in Developing Seeds. Arabidopsis Protein Disulfide Isomerase-5 Inhibits Cysteine Proteases during Trafficking to Vacuoles before Programmed Cell Death of the Endothelium in Developing Seeds. Plant Cell (1040-4651) 2008 imex curation Andeme Ondzighi C., Christopher DA., Cho EJ., Chang SC., Staehelin LA. Insufficient information was provided by the authors to unambigiously identify the positive control proteins used in this study. ondzighi@colorado.edu 16-Oct-2008: Contacted by JYOTI. jyoti Only protein-protein interactions 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 Andeme Ondzighi C., Christopher DA., Cho EJ., Chang SC., Staehelin LA. Insufficient information was provided by the authors to unambigiously identify the positive control proteins used in this study. ondzighi@colorado.edu Accepted 2008-OCT-22 by JYOTI. 16-Oct-2008: Contacted by JYOTI. Plant Cell (1040-4651) 2008 jyoti Only protein-protein interactions imex curation Arabidopsis Protein Disulfide Isomerase-5 Inhibits Cysteine Proteases during Trafficking to Vacuoles before Programmed Cell Death of the Endothelium in Developing Seeds. Arabidopsis Protein Disulfide Isomerase-5 Inhibits Cysteine Proteases during Trafficking to Vacuoles before Programmed Cell Death of the Endothelium in Developing Seeds. Plant Cell (1040-4651) 2008 imex curation Andeme Ondzighi C., Christopher DA., Cho EJ., Chang SC., Staehelin LA. Insufficient information was provided by the authors to unambigiously identify the positive control proteins used in this study. ondzighi@colorado.edu 16-Oct-2008: Contacted by JYOTI. jyoti Only protein-protein interactions in vitro In vitro anti bait coip anti bait coimmunoprecipitation anti bait coip anti tag western anti tag western blot anti tag western Andeme Ondzighi C., Christopher DA., Cho EJ., Chang SC., Staehelin LA. Insufficient information was provided by the authors to unambigiously identify the positive control proteins used in this study. ondzighi@colorado.edu Accepted 2008-OCT-22 by JYOTI. 16-Oct-2008: Contacted by JYOTI. Plant Cell (1040-4651) 2008 jyoti Only protein-protein interactions imex curation Arabidopsis Protein Disulfide Isomerase-5 Inhibits Cysteine Proteases during Trafficking to Vacuoles before Programmed Cell Death of the Endothelium in Developing Seeds. Arabidopsis Protein Disulfide Isomerase-5 Inhibits Cysteine Proteases during Trafficking to Vacuoles before Programmed Cell Death of the Endothelium in Developing Seeds. Plant Cell (1040-4651) 2008 imex curation Andeme Ondzighi C., Christopher DA., Cho EJ., Chang SC., Staehelin LA. Insufficient information was provided by the authors to unambigiously identify the positive control proteins used in this study. ondzighi@colorado.edu 16-Oct-2008: Contacted by JYOTI. jyoti Only protein-protein interactions 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 Andeme Ondzighi C., Christopher DA., Cho EJ., Chang SC., Staehelin LA. Insufficient information was provided by the authors to unambigiously identify the positive control proteins used in this study. ondzighi@colorado.edu Accepted 2008-OCT-22 by JYOTI. 16-Oct-2008: Contacted by JYOTI. Arabidopsis 3 day old etiolated seedling cDNA library and Arabidopsis 3 day old seedling treated with osmotic and hormone treatments. Plant Cell (1040-4651) 2008 jyoti Only protein-protein interactions imex curation Arabidopsis Protein Disulfide Isomerase-5 Inhibits Cysteine Proteases during Trafficking to Vacuoles before Programmed Cell Death of the Endothelium in Developing Seeds. Arabidopsis Protein Disulfide Isomerase-5 Inhibits Cysteine Proteases during Trafficking to Vacuoles before Programmed Cell Death of the Endothelium in Developing Seeds. Plant Cell (1040-4651) 2008 imex curation Andeme Ondzighi C., Christopher DA., Cho EJ., Chang SC., Staehelin LA. Insufficient information was provided by the authors to unambigiously identify the positive control proteins used in this study. ondzighi@colorado.edu 16-Oct-2008: Contacted by JYOTI. jyoti Only protein-protein interactions arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress electron microscopy electron microscopy Electron crystallography Electron cryomicroscopy Electron crystallography Electron cryomicroscopy immunostaining immunostaining Andeme Ondzighi C., Christopher DA., Cho EJ., Chang SC., Staehelin LA. Insufficient information was provided by the authors to unambigiously identify the positive control proteins used in this study. ondzighi@colorado.edu Accepted 2008-OCT-22 by JYOTI. 16-Oct-2008: Contacted by JYOTI. Plant Cell (1040-4651) 2008 jyoti Only protein-protein interactions imex curation rd21c_arath Probable cysteine protease RD21C RD21C MLD14.12 At3g19390 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MATSIKSITLALLIFSVLLISLSLGSVTATETTRNEAEARRMYERWLVENRKNYNGLGEKERRFEIFKDNLKFVEEHSSIPNRTYEVGLTRFADLTNDEFRAIYLRSKMERTRVPVKGEKYLYKVGDSLPDAIDWRAKGAVNPVKDQGSCGSCWAFSAIGAVEGINQIKTGELISLSEQELVDCDTSYNDGCGGGLMDYAFKFIIENGGIDTEEDYPYIATDVNVCNSDKKNTRVVTIDGYEDVPQNDEKSLKKALANQPISVAIEAGGRAFQLYTSGVFTGTCGTSLDHGVVAVGYGSEGGQDYWIVRNSWGSNWGESGYFKLERNIKESSGKCGVAMMASYPTKSSGSNPPKPPAPSPVVCDKSNTCPAKSTCCCLYEYNGKCYSWGCCPYESATCCDDGSSCCPQSYPVCDLKANTCRMKGNSPLSIKALTRGPAIATTKSTNMLVGSA CF1D2CAA957FC429 rd21b_arath Probable cysteine protease RD21B RD21B MMG4.7 At5g43060 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MGFLKLSPMILLLAMIGVSYAMDMSIISYDENHHITTETSRSDSEVERIYEAWMVEHGKKKMNQNGLGAEKDQRFEIFKDNLRFIDEHNTKNLSYKLGLTRFADLTNEEYRSMYLGAKPTKRVLKTSDRYQARVGDALPDSVDWRKEGAVADVKDQGSCGSCWAFSTIGAVEGINKIVTGDLISLSEQELVDCDTSYNQGCNGGLMDYAFEFIIKNGGIDTEADYPYKAADGRCDQNRKNAKVVTIDSYEDVPENSEASLKKALAHQPISVAIEAGGRAFQLYSSGVFDGLCGTELDHGVVAVGYGTENGKDYWIVRNSWGNRWGESGYIKMARNIEAPTGKCGIAMEASYPIKKGQNPPNPGPSPPSPIKPPTTCDKYFSCPESNTCCCLYKYGKYCFGWGCCPLEAATCCDDNSSCCPHEYPVCDVNRGTCLMSKNSPFSVKALKRTPAIPFWAKSRKHIA 699E9BC9B42A3652 pdi11_arath Protein disulfide isomerase-like 1-1 At1g21750 F8K7.19 PDIL1-1 PDI5 Protein disulfide-isomerase 1 Protein disulfide-isomerase 5 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MAMRGFTLFSILVLSLCASSIRSEETETKEFVLTLDHTNFTDTINKHDFIVVEFYAPWCGHCKQLAPEYEKAASALSSNVPPVVLAKIDASEETNREFATQYEVQGFPTIKIFRNGGKAVQEYNGPREAEGIVTYLKKQSGPASAEIKSADDASEVVSDKKVVVVGIFPKLSGSEFDSFMAIAEKLRSELDFAHTSDAKLLPRGESSVTGPVVRLFKPFDEQFVDSKDFDGEALEKFVKESSIPLITVFDKDPNNHPYVIKFFESTNTKAMLFINFTGEGAESLKSKYREVATSNKGQGLSFLLGDAENSQGAFQYFGLEESQVPLIIIQTADDKKYLKTNVEVDQIESWVKDFKDGKIAPHKKSQPIPAENNEPVKVVVSDSLDDIVLNSGKNVLLEFYAPWCGHCQKLAPILDEVAVSYQSDSSVVIAKLDATANDFPKDTFDVKGFPTIYFKSASGNVVVYEGDRTKEDFISFVDKNKDTVGEPKKEEETTEEVKDEL C27C564D38ECA6E8 rd21a_arath Cysteine proteinase RD21A RD21A Protein RESPONSIVE TO DEHYDRATION 21 F2G19.31 At1g47128 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MGFLKPTMAILFLAMVAVSSAVDMSIISYDEKHGVSTTGGRSEAEVMSIYEAWLVKHGKAQSQNSLVEKDRRFEIFKDNLRFVDEHNEKNLSYRLGLTRFADLTNDEYRSKYLGAKMEKKGERRTSLRYEARVGDELPESIDWRKKGAVAEVKDQGGCGSCWAFSTIGAVEGINQIVTGDLITLSEQELVDCDTSYNEGCNGGLMDYAFEFIIKNGGIDTDKDYPYKGVDGTCDQIRKNAKVVTIDSYEDVPTYSEESLKKAVAHQPISIAIEAGGRAFQLYDSGIFDGSCGTQLDHGVVAVGYGTENGKDYWIVRNSWGKSWGESGYLRMARNIASSSGKCGIAIEPSYPIKNGENPPNPGPSPPSPIKPPTQCDSYYTCPESNTCCCLFEYGKYCFAWGCCPLEAATCCDDNYSCCPHEYPVCDLDQGTCLLSKNSPFSVKALKRKPATPFWSQGRKNIA 4C59955CCB95AA58 at1g21750-rd21a 3 8 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association To elucidate the function of PDI5 in endothelial cell vacuoles, we screened two Arabidopsis cDNA libraries for proteins interacting with PDI5 bait using the yeast two-hybrid system. The principal isolates were clones encoding the following three Cys proteases: RD21 (NM_103612),CP43 (NM_123672), andCP19 (NM_112826), which represented 66, 31, and 3.8% of the total recombinants isolated, respectively. at1g21750-at5g43060 3 7 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 6 unspecified role unspecified role prey prey physical association physical association To elucidate the function of PDI5 in endothelial cell vacuoles, we screened two Arabidopsis cDNA libraries for proteins interacting with PDI5 bait using the yeast two-hybrid system. The principal isolates were clones encoding the following three Cys proteases: RD21 (NM_103612),CP43 (NM_123672), andCP19 (NM_112826), which represented 66, 31, and 3.8% of the total recombinants isolated, respectively. at1g21750-at3g19390 3 7 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 5 unspecified role unspecified role prey prey physical association physical association To elucidate the function of PDI5 in endothelial cell vacuoles, we screened two Arabidopsis cDNA libraries for proteins interacting with PDI5 bait using the yeast two-hybrid system. The principal isolates were clones encoding the following three Cys proteases: RD21 (NM_103612),CP43 (NM_123672), andCP19 (NM_112826), which represented 66, 31, and 3.8% of the total recombinants isolated, respectively. at1g21750-at3g1939-1 1 7 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 5 unspecified role unspecified role prey prey physical association physical association To elucidate the function of PDI5 in endothelial cell vacuoles, we screened two Arabidopsis cDNA libraries for proteins interacting with PDI5 bait using the yeast two-hybrid system. The principal isolates were clones encoding the following three Cys proteases: RD21 (NM_103612),CP43 (NM_123672), andCP19 (NM_112826), which represented 66, 31, and 3.8% of the total recombinants isolated, respectively. Figure 7A Figure 7B at1g21750-at5g4306-1 1 7 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 6 unspecified role unspecified role prey prey physical association physical association To elucidate the function of PDI5 in endothelial cell vacuoles, we screened two Arabidopsis cDNA libraries for proteins interacting with PDI5 bait using the yeast two-hybrid system. The principal isolates were clones encoding the following three Cys proteases: RD21 (NM_103612),CP43 (NM_123672), andCP19 (NM_112826), which represented 66, 31, and 3.8% of the total recombinants isolated, respectively. Figure 7A Figure 7B at1g21750-rd21a-1 1 7 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain 8 unspecified role unspecified role prey prey physical association physical association To elucidate the function of PDI5 in endothelial cell vacuoles, we screened two Arabidopsis cDNA libraries for proteins interacting with PDI5 bait using the yeast two-hybrid system. The principal isolates were clones encoding the following three Cys proteases: RD21 (NM_103612),CP43 (NM_123672), andCP19 (NM_112826), which represented 66, 31, and 3.8% of the total recombinants isolated, respectively. Figure 7A Figure 7B at1g21750-rd21a-2 2 7 unspecified role unspecified role bait bait ecoli Escherichia coli (strain K12) 8 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 ecoli Escherichia coli (strain K12) physical association physical association We conducted coimmunoprecipitation assays on the two most abundant recombinant Cys proteases (RD21 and CP43) isolated (Figure 7C). The PDI5 antiserumcoimmunoprecipitatedwithRD21 and CP43 only in the presence of recombinant PDI5 Figure 7C at1g21750-at5g4306-2 2 6 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 ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role bait bait ecoli Escherichia coli (strain K12) direct interaction direct interaction We conducted coimmunoprecipitation assays on the two most abundant recombinant Cys proteases (RD21 and CP43) isolated (Figure 7C). The PDI5 antiserumcoimmunoprecipitatedwithRD21 and CP43 only in the presence of recombinant PDI5 Figure 7C at1g21750-rd21a-3 4 7 unspecified role unspecified role neutral component neutral component 8 unspecified role unspecified role neutral component neutral component colocalization colocalization PDI5 and RD21 colocalized in ER cisternae (Figures 8A and 8D), LVs (Figure 8B),PSVs (Figure 8C), the Golgi, trans-Golgi network (TGN), and vesicles (Figure 8D) of endothelial cells. Figure 8