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 Snf2 homolog Fun30 acts as a homodimeric ATP-dependent chromatin-remodeling enzyme. The Snf2 homolog Fun30 acts as a homodimeric ATP-dependent chromatin-remodeling enzyme. J. Biol. Chem. (0021-9258) 2010 imex curation Awad S., Ryan D., Prochasson P., Owen-Hughes T., Hassan A.H. ahmedh@uaeu.ac.ae 05-Jan-2015: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Awad S., Ryan D., Prochasson P., Owen-Hughes T., Hassan A.H. ahmedh@uaeu.ac.ae Accepted 2014-DEC-12 by MGT J. Biol. Chem. (0021-9258) 2010 Only protein-protein interactions imex curation The Snf2 homolog Fun30 acts as a homodimeric ATP-dependent chromatin-remodeling enzyme. The Snf2 homolog Fun30 acts as a homodimeric ATP-dependent chromatin-remodeling enzyme. J. Biol. Chem. (0021-9258) 2010 imex curation Awad S., Ryan D., Prochasson P., Owen-Hughes T., Hassan A.H. ahmedh@uaeu.ac.ae 05-Jan-2015: Contacted by IntAct-Help. Only protein-protein interactions yeast Saccharomyces cerevisiae Baker's yeast anti tag coip anti tag coimmunoprecipitation anti tag coip anti tag western anti tag western blot anti tag western Awad S., Ryan D., Prochasson P., Owen-Hughes T., Hassan A.H. ahmedh@uaeu.ac.ae Accepted 2014-DEC-12 by MGT J. Biol. Chem. (0021-9258) 2010 Only protein-protein interactions imex curation The Snf2 homolog Fun30 acts as a homodimeric ATP-dependent chromatin-remodeling enzyme. The Snf2 homolog Fun30 acts as a homodimeric ATP-dependent chromatin-remodeling enzyme. J. Biol. Chem. (0021-9258) 2010 imex curation Awad S., Ryan D., Prochasson P., Owen-Hughes T., Hassan A.H. ahmedh@uaeu.ac.ae 05-Jan-2015: Contacted by IntAct-Help. Only protein-protein interactions yeast Saccharomyces cerevisiae 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 Awad S., Ryan D., Prochasson P., Owen-Hughes T., Hassan A.H. The complex was first purified by TAP and shown to be pure by silver staining. ahmedh@uaeu.ac.ae Accepted 2014-DEC-12 by MGT J. Biol. Chem. (0021-9258) 2010 Only protein-protein interactions imex curation The Snf2 homolog Fun30 acts as a homodimeric ATP-dependent chromatin-remodeling enzyme. The Snf2 homolog Fun30 acts as a homodimeric ATP-dependent chromatin-remodeling enzyme. J. Biol. Chem. (0021-9258) 2010 imex curation Awad S., Ryan D., Prochasson P., Owen-Hughes T., Hassan A.H. ahmedh@uaeu.ac.ae 05-Jan-2015: Contacted by IntAct-Help. Only protein-protein interactions yeast Saccharomyces cerevisiae Baker's yeast density sedimentation cosedimentation through density gradient density sedimentation weight by comassie molecular weight estimation by coomasie staining weight by comassie Awad S., Ryan D., Prochasson P., Owen-Hughes T., Hassan A.H. The complex was first purified by TAP and shown to be pure by silver staining. Fun30 was resolved by 10–40% glycerol gradient sedimentation together with amylase and apoferritin as molecular weight markers. ahmedh@uaeu.ac.ae Accepted 2014-DEC-12 by MGT J. Biol. Chem. (0021-9258) 2010 Only protein-protein interactions imex curation The Snf2 homolog Fun30 acts as a homodimeric ATP-dependent chromatin-remodeling enzyme. The Snf2 homolog Fun30 acts as a homodimeric ATP-dependent chromatin-remodeling enzyme. J. Biol. Chem. (0021-9258) 2010 imex curation Awad S., Ryan D., Prochasson P., Owen-Hughes T., Hassan A.H. ahmedh@uaeu.ac.ae 05-Jan-2015: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro atpase assay atpase assay predetermined predetermined participant predetermined Awad S., Ryan D., Prochasson P., Owen-Hughes T., Hassan A.H. ahmedh@uaeu.ac.ae Accepted 2014-DEC-12 by MGT J. Biol. Chem. (0021-9258) 2010 Only protein-protein interactions imex curation atp ATP Adenosine 5'-triphosphate Adenosine triphosphate ADENOSINE-5'-TRIPHOSPHATE Adephos Adetol Adynol Atipi ATP Atriphos Cardenosine Fosfobion Glucobasin H4atp Myotriphos Triadenyl Triphosphaden adenosine 5'-(tetrahydrogen triphosphate) small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) fun30_yeast ATP-dependent helicase FUN30 YAL001 YAL019W FUN30 protein protein yeast Saccharomyces cerevisiae Baker's yeast MSGSHSNDEDDVVQVPETSSPTKVASSSPLKPTSPTVPDASVASLRSRFTFKPSDPSEGAHTSKPLPSGSPEVALVNLAREFPDFSQTLVQAVFKSNSFNLQSARERLTRLRQQRQNWTWNKNASPKKSETPPPVKKSLPLANTGRLSSIHGNINNKSSKITVAKQKTSIFDRYSNVINQKQYTFELPTNLNIDSEALSKLPVNYNKKRRLVRADQHPIGKSYESSATQLGSAREKLLANRKYGRHANDNDEEEEESMMTDDDDASGDDYTESTPQINLDEQVLQFINDSDIVDLSDLSDTTMHKAQLIASHRPYSSLNAFVNTNFNDKDTEENASNKRKRRAAASANESERLLDKITQSIRGYNAIESVIKKCSSYGDLVTSQMKKWGVQVEGDNSELDLMNLGEDDDDDNDDGNNDNNNSNNNNTAGADATSKEKEDTKAVVEGFDETSAEPTPAPAPAPVERETKRIRNTTKPKVVEDEDDDVDLEAIDDELPQSEHEDDDYEEEDEDYNDEEEDVEYDDGDDDDDDDDEFVATRKNTHVISTTSRNGRKPIVKFFKGKPRLLSPEISLKDYQQTGINWLNLLYQNKMSCILADDMGLGKTCQVISFFAYLKQINEPGPHLVVVPSSTLENWLREFQKFAPALKIEPYYGSLQEREELRDILERNAGKYDVIVTTYNLAAGNKYDVSFLKNRNFNVVVYDEGHMLKNSTSERFAKLMKIRANFRLLLTGTPLQNNLKELMSLLEFIMPNLFISKKESFDAIFKQRAKTTDDNKNHNPLLAQEAITRAKTMMKPFILRRRKDQVLKHLPPKHTHIQYCELNAIQKKIYDKEIQIVLEHKRMIKDGELPKDAKEKSKLQSSSSKNLIMALRKASLHPLLFRNIYNDKIITKMSDAILDEPAYAENGNKEYIKEDMSYMTDFELHKLCCNFPNTLSKYQLHNDEWMQSGKIDALKKLLKTIIVDKQEKVLIFSLFTQVLDILEMVLSTLDYKFLRLDGSTQVNDRQLLIDKFYEDKDIPIFILSTKAGGFGINLVCANNVIIFDQSFNPHDDRQAADRAHRVGQTKEVNITTLITKDSIEEKIHQLAKNKLALDSYISEDKKSQDVLESKVSDMLEDIIYDENSKPKGTKE 7E61B9ABB3A42ED2 7E61B9ABB3A42ED2 fun30-1 3 7 unspecified role unspecified role neutral component neutral component region tap tagged calmodulin binding peptide plus protein a tag CBP-ProtA tagged CBP-ProtA tagged tap tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Fig. 1B fun30-1 4 7 unspecified role unspecified role neutral component neutral component region tap tagged calmodulin binding peptide plus protein a tag CBP-ProtA tagged CBP-ProtA tagged tap tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Fig. 1C fun30-fun30 2 7 unspecified role unspecified role bait bait region tap tagged calmodulin binding peptide plus protein a tag CBP-ProtA tagged CBP-ProtA tagged tap tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 7 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 physical association physical association Fig. 2A fun30-fun30-1 2 7 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 7 unspecified role unspecified role prey prey region tap tagged calmodulin binding peptide plus protein a tag CBP-ProtA tagged CBP-ProtA tagged tap tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association Fig. 2A fun30-fun30-3 1 7 unspecified role unspecified role prey prey region tap tagged calmodulin binding peptide plus protein a tag CBP-ProtA tagged CBP-ProtA tagged tap tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yeast Saccharomyces cerevisiae Baker's yeast 7 unspecified role unspecified role bait bait c-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range ecoli Escherichia coli (strain K12) physical association physical association Fig. 2B atp-fun30 5 7 enzyme enzyme unspecified role unspecified role region tap tagged calmodulin binding peptide plus protein a tag CBP-ProtA tagged CBP-ProtA tagged tap tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined yeast Saccharomyces cerevisiae Baker's yeast 6 enzyme target enzyme target substrate substrate unspecified role unspecified role region 32p radiolabel 32p radiolabel P32 32P 32P P32 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined atpase reaction atpase reaction DNA or chromatin were present Fig.3