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/ chen-1999a-1 The spindle checkpoint of budding yeast depends on a tight complex between the Mad1 and Mad2 proteins. yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Chen RH., Brady DM., Smith D., Murray AW., Hardwick KG. rc70@cornell.edu Mol. Biol. Cell (1059-1524) 1999 Only protein-protein interactions imex curation chen-1999a-2 The spindle checkpoint of budding yeast depends on a tight complex between the Mad1 and Mad2 proteins. yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column western blot western blot Immuno blot Immuno blot Chen RH., Brady DM., Smith D., Murray AW., Hardwick KG. Superose 6 fast performance liquid chromatography column rc70@cornell.edu Mol. Biol. Cell (1059-1524) 1999 Only protein-protein interactions imex curation chen-1999a-3 The spindle checkpoint of budding yeast depends on a tight complex between the Mad1 and Mad2 proteins. chlae-cos_7 Cercopithecus aethiops simian cells transformed with SV40 Grivet cos_7 cercopithecus aethiops, by transformation with SV40 anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Chen RH., Brady DM., Smith D., Murray AW., Hardwick KG. rc70@cornell.edu Mol. Biol. Cell (1059-1524) 1999 Only protein-protein interactions imex curation chen-1999a-4 The spindle checkpoint of budding yeast depends on a tight complex between the Mad1 and Mad2 proteins. yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Chen RH., Brady DM., Smith D., Murray AW., Hardwick KG. rc70@cornell.edu Mol. Biol. Cell (1059-1524) 1999 Only protein-protein interactions imex curation mad1_yeast Spindle assembly checkpoint component MAD1 MAD1 Mitotic arrest deficient protein 1 YGL086W protein protein yeast Saccharomyces cerevisiae Baker's yeast MDVRAALQCFFSALSGRFTGKKLGLEIYSIQYKMSNSGGSSPFLESPGGSPDVGSTNGQSNRQIQALQFKLNTLQNEYEIEKLQLQKQTNILEKKYKATIDELEKALNDTKYLYESNDKLEQELKSLKERSANSMNDKDKCIEELRTTLQNKDLEMETLRQQYDSKLSKVTNQCDHFKLEAESSHSLLMKYEKEIKRQSVDIKDLQHQVMEKDDELSSVKASKMINSHPNYSTEEFNELTEMNKMIQDQVQYTKELELANMQQANELKKLKQSQDTSTFWKLENEKLQNKLSQLHVLESQYENLQLENIDLKSKLTKWEIYNDSDDDDDNNVNNNDNNNNNKNDNNNDNNNDTSNNNNINNNNRTKNNIRNNPEEIIRDWKLTKKECLILTDMNDKLRLDNNNLKLLNDEMALERNQILDLNKNYENNIVNLKRLNHELEQQKSLSFEECRLLREQLDGLYSAQNNALLEVENSETHASNKNVNEDMNNLIDTYKNKTEDLTNELKKLNDQLLSNSNDVETQRKKRKLTSDQIGLNYSQRLNELQLENVSVSRELSKAQTTIQLLQEKLEKLTKLKEKKIRILQLRDGPFIKDQFIKKNKLLLLEKENADLLNELKKNNPAVETVPISVYDSLNFELKQFEQEVFKSNKRFSRLKQVFNNKSLEFIDVVNSLLGFKLEFQQDSRVKIFSCFKPEKYLIADLNENTLKSNLDADIEGWDDLMNLWVEDRGQLPCFLATITLRLWEQRQAK mad2_yeast Mitotic spindle checkpoint component MAD2 MAD2 J1256 YJL030W protein protein yeast Saccharomyces cerevisiae Baker's yeast MSQSISLKGSTRTVTEFFEYSINSILYQRGVYPAEDFVTVKKYDLTLLKTHDDELKDYIRKILLQVHRWLLGGKCNQLVLCIVDKDEGEVVERWSFNVQHISGNSNGQDDVVDLNTTQSQIRALIRQITSSVTFLPELTKEGGYTFTVLAYTDADAKVPLEWADSNSKEIPDGEVVQFKTFSTNDHKVGAQVSYKY mad1-mad2-1 Interaction between Mad1 and Mad2 identified by coimunoprecipitation 653252 N/A 653256 unspecified role unspecified role prey prey c-terminal 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 653257 unspecified role unspecified role bait bait c-terminal required to bind necessary binding region required to bind deletion analysis deletion analysis undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false 653262 653260 physical association physical association false false false Figure 3A, 4A, 8A Interaction was shown to be independent of cell cycle or phosphorylation state of Mad1 mad1-mad2-2 Interaction between Mad1 and Mad2identified by gel filtration 653253 N/A 653256 unspecified role unspecified role neutral component neutral component N/A 653257 unspecified role unspecified role neutral component neutral component physical association physical association false false false Figure 3B MW = greater than 670 kDa. mad1-mad2-3 Interaction between Mad1 and Mad2 identified by coimunoprecipitation 653254 N/A 653256 unspecified role unspecified role prey prey N/A 653257 unspecified role unspecified role bait bait physical association physical association false false false Figure 6 Interaction was shown to be independent of the phosphorylation state of Mad1 mad1-mad2-4 Interaction between Mad1 and Mad2 identified by coimunoprecipitation 653255 N/A 653256 unspecified role unspecified role bait bait region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false gal4 dna_bd tag tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 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 653257 unspecified role unspecified role prey prey physical association physical association false false false Figure 7C