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/ RAC, a stable ribosome-associated complex in yeast formed by the DnaK-DnaJ homologs Ssz1p and zuotin. RAC, a stable ribosome-associated complex in yeast formed by the DnaK-DnaJ homologs Ssz1p and zuotin. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2001 imex curation Gautschi M., Lilie H., Funfschilling U., Mun A., Ross S., Lithgow T., Rucknagel P., Rospert S. Proteins were released from ribosomes by treatment with 700 mM Kacetate, then sequentially purified on ResourceQ, Superdex200 and MonoQ. rospert@enzyme-halle.mpg.de 23-Feb-2010: Contacted by JYOTI. jyoti Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast solution sedimentati cosedimentation in solution solution sedimentati western blot western blot Immuno blot Immuno blot Gautschi M., Lilie H., Funfschilling U., Mun A., Ross S., Lithgow T., Rucknagel P., Rospert S. Proteins were released from ribosomes by treatment with 700 mM Kacetate, then sequentially purified on ResourceQ, Superdex200 and MonoQ. rospert@enzyme-halle.mpg.de Accepted 2009-Not a month!-23 by JYOTI. 23-Feb-2010: Contacted by JYOTI. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2001 jyoti Only protein-protein interactions imex curation RAC, a stable ribosome-associated complex in yeast formed by the DnaK-DnaJ homologs Ssz1p and zuotin. RAC, a stable ribosome-associated complex in yeast formed by the DnaK-DnaJ homologs Ssz1p and zuotin. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2001 imex curation Gautschi M., Lilie H., Funfschilling U., Mun A., Ross S., Lithgow T., Rucknagel P., Rospert S. Proteins were released from ribosomes by treatment with 700 mM Kacetate, then sequentially purified on ResourceQ, Superdex200 and MonoQ. rospert@enzyme-halle.mpg.de 23-Feb-2010: Contacted by JYOTI. jyoti Only protein-protein interactions 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 Gautschi M., Lilie H., Funfschilling U., Mun A., Ross S., Lithgow T., Rucknagel P., Rospert S. Proteins were released from ribosomes by treatment with 700 mM Kacetate, then sequentially purified on ResourceQ, Superdex200 and MonoQ. rospert@enzyme-halle.mpg.de Accepted 2009-Not a month!-23 by JYOTI. 23-Feb-2010: Contacted by JYOTI. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2001 jyoti Only protein-protein interactions imex curation RAC, a stable ribosome-associated complex in yeast formed by the DnaK-DnaJ homologs Ssz1p and zuotin. RAC, a stable ribosome-associated complex in yeast formed by the DnaK-DnaJ homologs Ssz1p and zuotin. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2001 imex curation Gautschi M., Lilie H., Funfschilling U., Mun A., Ross S., Lithgow T., Rucknagel P., Rospert S. Proteins were released from ribosomes by treatment with 700 mM Kacetate, then sequentially purified on ResourceQ, Superdex200 and MonoQ. rospert@enzyme-halle.mpg.de 23-Feb-2010: Contacted by JYOTI. jyoti Only protein-protein interactions yeasx Saccharomyces cerevisiae (Baker's yeast) Baker's yeast chromatography chromatography technology column chromatography chromatography western blot western blot Immuno blot Immuno blot Gautschi M., Lilie H., Funfschilling U., Mun A., Ross S., Lithgow T., Rucknagel P., Rospert S. Proteins were released from ribosomes by treatment with 700 mM Kacetate, then sequentially purified on ResourceQ, Superdex200 and MonoQ. rospert@enzyme-halle.mpg.de Accepted 2009-Not a month!-23 by JYOTI. 23-Feb-2010: Contacted by JYOTI. Proc. Natl. Acad. Sci. U.S.A. (0027-8424) 2001 jyoti Only protein-protein interactions imex curation zuo1_yeast Zuotin Ribosome-associated complex subunit ZUO1 DnaJ-related protein ZUO1 Heat shock protein 40 homolog ZUO1 ZUO1 YGR285C protein protein yeast Saccharomyces cerevisiae Baker's yeast MFSLPTLTSDITVEVNSSATKTPFVRRPVEPVGKFFLQHAQRTLRNHTWSEFERIEAEKNVKTVDESNVDPDELLFDTELADEDLLTHDARDWKTADLYAAMGLSKLRFRATESQIIKAHRKQVVKYHPDKQSAAGGSLDQDGFFKIIQKAFETLTDSNKRAQYDSCDFVADVPPPKKGTDYDFYEAWGPVFEAEARFSKKTPIPSLGNKDSSKKEVEQFYAFWHRFDSWRTFEFLDEDVPDDSSNRDHKRYIERKNKAARDKKKTADNARLVKLVERAVSEDPRIKMFKEEEKKEKERRKWEREAGARAEAEAKAKAEAEAKAKAESEAKANASAKADKKKAKEAAKAAKKKNKRAIRNSAKEADYFGDADKATTIDEQVGLIVDSLNDEELVSTADKIKANAAGAKEVLKESAKTIVDSGKLPSSLLSYFV 0AA76BC11D3C7DAB 0AA76BC11D3C7DAB ssz1_yeast Ribosome-associated complex subunit SSZ1 YHR064C SSZ1 PDR13 Heat shock protein 70 homolog SSZ1 DnaK-related protein SSZ1 Pleiotropic drug resistance protein 13 protein protein yeast Saccharomyces cerevisiae Baker's yeast MSSPVIGITFGNTSSSIAYINPKNDVDVIANPDGERAIPSALSYVGEDEYHGGQALQQLIRNPKNTIINFRDFIGLPFDKCDVSKCANGAPAVEVDGKVGFVISRGEGKEEKLTVDEVVSRHLNRLKLAAEDYIGSAVKEAVLTVPTNFSEEQKTALKASAAKIGLQIVQFINEPSAALLAHAEQFPFEKDVNVVVADFGGIRSDAAVIAVRNGIFTILATAHDLSLGGDNLDTELVEYFASEFQKKYQANPRKNARSLAKLKANSSITKKTLSNATSATISIDSLADGFDYHASINRMRYELVANKVFAQFSSFVDSVIAKAELDPLDIDAVLLTGGVSFTPKLTTNLEYTLPESVEILGPQNKNASNNPNELAASGAALQARLISDYDADELAEALQPVIVNTPHLKKPIGLIGAKGEFHPVLLAETSFPVQKKLTLKQAKGDFLIGVYEGDHHIEEKTLEPIPKEENAEEDDESEWSDDEPEVVREKLYTLGTKLMELGIKNANGVEIIFNINKDGALRVTARDLKTGNAVKGEL 07B1A409B213E0B4 07B1A409B213E0B4 ssz1-zuo1-1 3 4 unspecified role unspecified role neutral component neutral component 5 unspecified role unspecified role neutral component neutral component physical association physical association The ratio between Ssz1p and zuotin was constant during purification, suggesting that the two proteins formed a ribosome-associated complex Fig. 1B ssz1-zuo1-2 2 4 unspecified role unspecified role prey prey 5 unspecified role unspecified role bait bait physical association physical association IgGs specific for either Ssz1p or zuotin immunoprecipitated RAC efficiently from the protein mixture released by salt wash from ribosomes. Fig. 3A Coimmunoprecipitation of the two subunits was unaffected by the addition of 2 mM ATP or ADP. A small amount of Ssa1/2p but was discounted by the author as non-specific binding. ssz1-zuo1-3 2 5 unspecified role unspecified role prey prey 4 unspecified role unspecified role bait bait physical association physical association IgGs specific for either Ssz1p or zuotin immunoprecipitated RAC efficiently from the protein mixture released by salt wash from ribosomes. Fig. 3A Coimmunoprecipitation of the two subunits was unaffected by the addition of 2 mM ATP or ADP. A small amount of Ssa1/2p but was discounted by the author as non-specific binding. ssz1-zuo1-4 1 4 unspecified role unspecified role neutral component neutral component 5 unspecified role unspecified role neutral component neutral component physical association physical association The stoichiometry between Ssz1p and zuotin in the complex was determined by analytical ultracentrifugation. Fig. 3B Complex m.w.=126,000 Da