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/ The mad2 conformational dimer: structure and implications for the spindle assembly checkpoint. The mad2 conformational dimer: structure and implications for the spindle assembly checkpoint. Cell (0092-8674) 2007 imex curation Mapelli M., Massimiliano L., Santaguida S., Musacchio A. andrea.musacchio@ifom-ieo-campus.it 01-April-2008: Contacted by JYOTI. Only protein-protein interactions 5 in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column predetermined predetermined participant predetermined Mapelli M., Massimiliano L., Santaguida S., Musacchio A. andrea.musacchio@ifom-ieo-campus.it Accepted 2008-FEB-31 by CDEROW. 01-April-2008: Contacted by JYOTI. Cell (0092-8674) 2007 Only protein-protein interactions 5 imex curation The mad2 conformational dimer: structure and implications for the spindle assembly checkpoint. The mad2 conformational dimer: structure and implications for the spindle assembly checkpoint. Cell (0092-8674) 2007 imex curation Mapelli M., Massimiliano L., Santaguida S., Musacchio A. andrea.musacchio@ifom-ieo-campus.it 01-April-2008: Contacted by JYOTI. Only protein-protein interactions 5 in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Mapelli M., Massimiliano L., Santaguida S., Musacchio A. andrea.musacchio@ifom-ieo-campus.it Accepted 2008-FEB-31 by CDEROW. 01-April-2008: Contacted by JYOTI. Cell (0092-8674) 2007 Only protein-protein interactions 5 imex curation The mad2 conformational dimer: structure and implications for the spindle assembly checkpoint. The mad2 conformational dimer: structure and implications for the spindle assembly checkpoint. Cell (0092-8674) 2007 imex curation Mapelli M., Massimiliano L., Santaguida S., Musacchio A. andrea.musacchio@ifom-ieo-campus.it 01-April-2008: Contacted by JYOTI. Only protein-protein interactions 5 in vitro In vitro pull down pull down predetermined predetermined participant predetermined Mapelli M., Massimiliano L., Santaguida S., Musacchio A. andrea.musacchio@ifom-ieo-campus.it Accepted 2008-FEB-31 by CDEROW. 01-April-2008: Contacted by JYOTI. Cell (0092-8674) 2007 Only protein-protein interactions 5 imex curation md2l1_human Mitotic spindle assembly checkpoint protein MAD2A MAD2L1 MAD2 Mitotic arrest deficient 2-like protein 1 protein protein human Homo sapiens Human MALQLSREQGITLRGSAEIVAEFFSFGINSILYQRGIYPSETFTRVQKYGLTLLVTTDLELIKYLNNVVEQLKDWLYKCSVQKLVVVISNIESGEVLERWQFDIECDKTAKDDSAPREKSQKAIQDEIRSVIRQITATVTFLPLLEVSCSFDLLIYTDKDLVVPEKWEESGPQFITNSEEVRLRSFTTTIHKVNSMVAYKIPVND DIP protein Q13257 original sequence version: 133 DIP protein Q13257 original sequence version: 78 DIP protein Q13257 original sequence version: 105 DIP protein Q13257 original sequence version: 120 DIP protein Q13257 original sequence version: 127 DIP protein Q13257 original sequence version: 116 DIP protein Q13257 original sequence version: 150 DIP protein Q13257 original sequence version: 156 DIP protein Q13257 original sequence version: 134 B8DCBF0043836764 mad2 peptide MAD2 binding peptide 1 peptide peptide polypeptide oligopeptide oligopeptide polypeptide in vitro In vitro SWYSYPPPQRAV An amino acid sequence which is not represented in UniProt 195CFA9B1877777B cdc20_human Cell division cycle protein 20 homolog CDC20 p55CDC protein protein human Homo sapiens Human MAQFAFESDLHSLLQLDAPIPNAPPARWQRKAKEAAGPAPSPMRAANRSHSAGRTPGRTPGKSSSKVQTTPSKPGGDRYIPHRSAAQMEVASFLLSKENQPENSQTPTKKEHQKAWALNLNGFDVEEAKILRLSGKPQNAPEGYQNRLKVLYSQKATPGSSRKTCRYIPSLPDRILDAPEIRNDYYLNLVDWSSGNVLAVALDNSVYLWSASSGDILQLLQMEQPGEYISSVAWIKEGNYLAVGTSSAEVQLWDVQQQKRLRNMTSHSARVGSLSWNSYILSSGSRSGHIHHHDVRVAEHHVATLSGHSQEVCGLRWAPDGRHLASGGNDNLVNVWPSAPGEGGWVPLQTFTQHQGAVKAVAWCPWQSNVLATGGGTSDRHIRIWNVCSGACLSAVDAHSQVCSILWSPHYKELISGHGFAQNQLVIWKYPTMAKVAELKGHTSRVLSLTMSPDGATVASAAADETLRLWRCFELDPARRREREKASAAKSSLIHQGIR DIP protein Q12834 original sequence version: 132 DIP protein Q12834 original sequence version: 74 DIP protein Q12834 original sequence version: 100 DIP protein Q12834 original sequence version: 118 DIP protein Q12834 original sequence version: 125 DIP protein Q12834 original sequence version: 114 DIP protein Q12834 original sequence version: 147 DIP protein Q12834 original sequence version: 162 FD5C967AF84089E8 mad2l1-mad2l1-1 1 5 unspecified role unspecified role neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) 4 unspecified role unspecified role neutral component neutral component mutated region TAKDDSAPR -> GSG experimental feature experimental feature substitut analysis substitution analysis substitut analysis certain certain sequence position certain certain certain sequence position certain ecobb Escherichia coli (strain B / BL21) 4 unspecified role unspecified role neutral component neutral component ecobb Escherichia coli (strain B / BL21) physical association physical association A stoichiometric combination of Mad2LL and Mad2wt-MBP1 generated a Mad2LL-Mad2wt-MBP1 complex eluting near the 44 kDa marker (Figure 1A). Figure 1A mad2l1-mad2l1-2 2 4 unspecified role unspecified role neutral component neutral component mutated region TAKDDSAPR -> GSG experimental feature experimental feature substitut analysis substitution analysis substitut analysis certain certain sequence position certain certain certain sequence position certain ecobb Escherichia coli (strain B / BL21) 4 unspecified role unspecified role neutral component neutral component region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecobb Escherichia coli (strain B / BL21) 5 unspecified role unspecified role neutral component neutral component chemical synthesis Chemical synthesis (Chemical synthesis) physical association physical association A stoichiometric combination of Mad2LL and Mad2wt-MBP1 generated a Mad2LL-Mad2wt-MBP1 complex eluting near the 44 kDa marker (Figure 1A). Table 1 We crystallized Mad2LL-Mad2w-MBP1 and collected X-ray diffraction data to 2.9 A resolution. 2.9 A Free = 27.4% cdc20-mad2l1-1 3 4 unspecified role unspecified role prey prey Q13257:p.Val193Gln mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain V Q Wed May 02 14:36:07 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'val193gln' Tue Jun 30 15:50:38 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Val193Gln' region required to bind necessary binding region required to bind certain certain sequence position certain certain certain sequence position certain ecobb Escherichia coli (strain B / BL21) 6 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 certain certain sequence position certain certain certain sequence position certain ecobb Escherichia coli (strain B / BL21) direct interaction direct interaction GST-Cdc20111-138 on GSH beads was incubated with the Mad2 species labeled in black on the upper part of the panel. Fig. 2F, 3B, 4D Loss of residues 196-205 force Mad2 into a closed conformation cdc20-mad2l1-2 3 4 unspecified role unspecified role prey prey ecobb Escherichia coli (strain B / BL21) 6 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 certain certain sequence position certain certain certain sequence position certain ecobb Escherichia coli (strain B / BL21) 4 unspecified role unspecified role prey prey region open conformation sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecobb Escherichia coli (strain B / BL21) physical association physical association GST-Cdc20111-138 on GSH beads was incubated with the Mad2 species labeled in black on the upper part of the panel. Figs. 2F, 4D, 5E Phe141 and Arg184 are required for the interaction of wild type Mad2 (C-MAD2) with Mad2 locked in the open conformation (O-MAD2), the same mutations in O-Mad2 did not significantly alter the binding to C-Mad2. Val193 generates a mutant that is locked as O-Mad2 and is totally defective in Cdc20 binding. Mutating Trp100 of Mad2 to Tyr (W100Y) generates a stable O-Mad2 mutant that binds to GST-Cdc20111�138 with very slow kinetics. mad2l1-mad2l1-3 1 4 unspecified role unspecified role neutral component neutral component Q13257:p.Leu13Ala closed conformation mutation increasing mutation increasing interaction mutation increasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain L A Wed May 02 16:49:40 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'leu13ala' Tue Jun 30 18:26:56 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Leu13Ala' ecobb Escherichia coli (strain B / BL21) 4 unspecified role unspecified role neutral component neutral component ecobb Escherichia coli (strain B / BL21) physical association physical association The shoulder in the elution peak of the Mad2L13A indicates that at 40 μM this mutant is a mixture of monomers and dimers. Supp. Fig 2