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/ A protein conjugation system essential for autophagy. A protein conjugation system essential for autophagy. Nature (0028-0836) 1998 imex curation Mizushima N., Noda T., Yoshimori T., Tanaka Y., Ishii T., George MD., Klionsky DJ., Ohsumi M., Ohsumi Y. yohsumi@nibb.ac.jp 25-Jun-2010: Contacted by JYOTI. jyoti Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Mizushima N., Noda T., Yoshimori T., Tanaka Y., Ishii T., George MD., Klionsky DJ., Ohsumi M., Ohsumi Y. yohsumi@nibb.ac.jp Accepted 2010-MAY-21 by JYOTI. Nature (0028-0836) 1998 jyoti Only protein-protein interactions imex curation A protein conjugation system essential for autophagy. A protein conjugation system essential for autophagy. Nature (0028-0836) 1998 imex curation Mizushima N., Noda T., Yoshimori T., Tanaka Y., Ishii T., George MD., Klionsky DJ., Ohsumi M., Ohsumi Y. yohsumi@nibb.ac.jp 25-Jun-2010: Contacted by JYOTI. jyoti Only protein-protein interactions in vitro In vitro density sedimentation cosedimentation through density gradient density sedimentation western blot western blot Immuno blot Immuno blot Mizushima N., Noda T., Yoshimori T., Tanaka Y., Ishii T., George MD., Klionsky DJ., Ohsumi M., Ohsumi Y. 10-step (18-54 % w/w) sucrose gradient. yohsumi@nibb.ac.jp Accepted 2010-MAY-21 by JYOTI. 25-Jun-2010: Contacted by JYOTI. Nature (0028-0836) 1998 jyoti Only protein-protein interactions imex curation atg5_yeast Autophagy protein 5 ATG5 APG5 YPL149W P2601 protein protein yeast Saccharomyces cerevisiae Baker's yeast MNDIKQLLWNGELNVLVSIDPSFLMKGSPREIAVLRIRVPRETYLVNYMPLIWNKIKSFLSFDPLTDSEKYFWFEHNKTPIPWNYPVGVLFDCLAGKSATFTTSFENQVKDVLTFLRIHLVMGDSLPPTIIPIASSKTQAEKFWFHQWKQVCFILNGSSKAIMSLSVNEARKFWGSVITRNFQDFIEISNKISSSRPRHIPLIIQTSRTSGTFRISQPTISMTGVNPTLKDIEGDILDVKEGINGNDVMVICQGIEIPWHMLLYDLYSKLRSFDGFLYITLVPIKGGDKASSEL BE513A631DEAA0A1 BE513A631DEAA0A1 atg12_yeast Ubiquitin-like protein ATG12 ATG12 APG12 YBR217W YBR1506 Autophagy-related protein 12 protein protein yeast Saccharomyces cerevisiae Baker's yeast MSRILESENETESDESSIISTNNGTAMERSRNNQELRSSPHTVQNRLELFSRRLSQLGLASDISVDQQVEDSSSGTYEQEETIKTNAQTSKQKSHKDEKNIQKIQIKFQPIGSIGQLKPSVCKISMSQSFAMVILFLKRRLKMDHVYCYINNSFAPSPQQNIGELWMQFKTNDELIVSYCASVAFG 74B3C9FD3CEA9265 74B3C9FD3CEA9265 atg5-atg12-1 1 3 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 4 unspecified role unspecified role prey prey region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Immunoprecipitation analysis revealed that the 70K band included both Apg5 and Apg12 (Fig. 2d). Fig. 2D atg5-atg12-2 2 3 unspecified role unspecified role neutral component neutral component region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast 4 unspecified role unspecified role neutral component neutral component region ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast physical association physical association We therefore examined their intracellular localization by sucrose density-gradient centrifugation analysis and found that free Apg5 and the Apg5/Apg12 conjugate co-fractionated (Fig. 6); in contrast, most of the Apg12 was in the denser fractions. Fig. 6B