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/ wang-2006b-1 Phosphorylation of HSF1 by MAPK-activated protein kinase 2 on serine 121, inhibits transcriptional activity and promotes HSP90 binding. human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Wang X., Khaleque MA., Zhao MJ., Zhong R., Gaestel M., Calderwood SK. BioCreative - Critical Assessment of Information Extraction systems in Biology scalderw@bidmc.harvard.edu J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation wang-2006b-2 Phosphorylation of HSF1 by MAPK-activated protein kinase 2 on serine 121, inhibits transcriptional activity and promotes HSP90 binding. in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Wang X., Khaleque MA., Zhao MJ., Zhong R., Gaestel M., Calderwood SK. BioCreative - Critical Assessment of Information Extraction systems in Biology scalderw@bidmc.harvard.edu J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation wang-2006b-3 Phosphorylation of HSF1 by MAPK-activated protein kinase 2 on serine 121, inhibits transcriptional activity and promotes HSP90 binding. in vitro In vitro pull down pull down ms participant Identification by mass spectrometry ms participant Wang X., Khaleque MA., Zhao MJ., Zhong R., Gaestel M., Calderwood SK. BioCreative - Critical Assessment of Information Extraction systems in Biology [32P]phosphopeptide fractions shown by the arrows in Figure 2 A are then analyzed by MALDI-TOF. scalderw@bidmc.harvard.edu J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation wang-2006b-4 Phosphorylation of HSF1 by MAPK-activated protein kinase 2 on serine 121, inhibits transcriptional activity and promotes HSP90 binding. human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column western blot western blot Immuno blot Immuno blot Wang X., Khaleque MA., Zhao MJ., Zhong R., Gaestel M., Calderwood SK. BioCreative - Critical Assessment of Information Extraction systems in Biology Whole cell extracts were applied to a Superdex 200 HR 10/30 column. scalderw@bidmc.harvard.edu J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation wang-2006b-5 Phosphorylation of HSF1 by MAPK-activated protein kinase 2 on serine 121, inhibits transcriptional activity and promotes HSP90 binding. human-hela Homo sapiens epitheloid cervix carcinoma cells Human hela Human epitheloid cervix carcinoma HeLa cells derivation: cervix; epithelial; adenocarcinoma ATCC no:CCL-2 anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Wang X., Khaleque MA., Zhao MJ., Zhong R., Gaestel M., Calderwood SK. BioCreative - Critical Assessment of Information Extraction systems in Biology scalderw@bidmc.harvard.edu J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation mapk2_human MAP kinase-activated protein kinase 2 MAPKAPK2 protein protein human Homo sapiens Human MLSNSQGQSPPVPFPAPAPPPQPPTPALPHPPAQPPPPPPQQFPQFHVKSGLQIKKNAIIDDYKVTSQVLGLGINGKVLQIFNKRTQEKFALKMLQDCPKARREVELHWRASQCPHIVRIVDVYENLYAGRKCLLIVMECLDGGELFSRIQDRGDQAFTEREASEIMKSIGEAIQYLHSINIAHRDVKPENLLYTSKRPNAILKLTDFGFAKETTSHNSLTTPCYTPYYVAPEVLGPEKYDKSCDMWSLGVIMYILLCGYPPFYSNHGLAISPGMKTRIRMGQYEFPNPEWSEVSEEVKMLIRNLLKTEPTQRMTITEFMNHPWIMQSTKVPQTPLHTSRVLKEDKERWEDVKEEMTSALATMRVDYEQIKIKKIEDASNPLLLKRRKKARALEAAALAH hsf1_human Heat shock factor protein 1 Heat shock transcription factor 1 HSF1 HSTF1 protein protein human Homo sapiens Human MDLPVGPGAAGPSNVPAFLTKLWTLVSDPDTDALICWSPSGNSFHVFDQGQFAKEVLPKYFKHNNMASFVRQLNMYGFRKVVHIEQGGLVKPERDDTEFQHPCFLRGQEQLLENIKRKVTSVSTLKSEDIKIRQDSVTKLLTDVQLMKGKQECMDSKLLAMKHENEALWREVASLRQKHAQQQKVVNKLIQFLISLVQSNRILGVKRKIPLMLNDSGSAHSMPKYSRQFSLEHVHGSGPYSAPSPAYSSSSLYAPDAVASSGPIISDITELAPASPMASPGGSIDERPLSSSPLVRVKEEPPSPPQSPRVEEASPGRPSSVDTLLSPTALIDSILRESEPAPASVTALTDARGHTDTEGRPPSPPPTSTPEKCLSVACLDKNELSDHLDAMDSNLDNLQTMLSSHGFSVDTSALLDLFSPSVTVPDMSLPDLDSSLASIQELLSPQEPPRPPEAENSSPDSGKQLVHYTAQPLFLLDPGSVDTGSNDLPVLFELGEGSYFSEGDGFAEDPTISLLTGSEPPKAKDPTVS hsf1-mk2 502112 N/A 502117 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 false thr334 opthr O-phospho-L-threonine (2S,3R)-2-amino-3-(phosphonooxy)butanoic acid 2-amino-3-hydroxybutanoic acid 3-phosphate 2-azanyl-3-(phosphonooxy)butanoic acid MOD_RES Phosphothreonine O-phospho-L-threonine O-phosphorylated L-threonine O3-phosphothreonine threonine phosphate ester OPThr Phospho Phospho Threonyl Phosphorylation phosphorylationt certain certain sequence position certain certain certain sequence position certain false N/A 502118 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 physical association physical association false false false 1C Decrease in Phosphothreonine resulted in decrease in interaction. hsf1-mk2-1 502113 N/A 502117 unspecified role unspecified role prey prey region 32p radiolabel 32p radiolabel P32 32P 32P P32 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 502118 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 false phosphorylation phosphorylation reaction phosphorylation false false false 2A, 2B Increase in labelled 32P was observed upto about 4 hrs. NaSal inhibited 32P incorporation GST-HSF1 was phosphorylated by MK2 hsf1-mk2-2 502114 N/A 502117 unspecified role unspecified role prey prey region 32p radiolabel 32p radiolabel P32 32P 32P P32 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false N/A 502118 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 false phosphorylation phosphorylation reaction phosphorylation false false false 2D, 2E HSF1 was phosphorylated. Sites phosphorylated 118-127 KVTSVSTLKS and 523-529 KAKDPTVS hsf1-mk2-3 502115 N/A 502118 unspecified role unspecified role neutral component neutral component N/A 502117 unspecified role unspecified role neutral component neutral component physical association physical association false false false HSP90 was also copurfied however whether the isoform A or B was purified could not be ascertained. 4A mk2-hsf1 502116 N/A 502117 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 false N/A 502118 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 Q00613:p.Ser121Ala mutation increasing mutation increasing interaction mutation increasing certain certain sequence position certain certain certain sequence position certain false Q00613:p.Ser121Asp mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing certain certain sequence position certain certain certain sequence position certain false physical association physical association false false false Enodgenous HSP90 was immunoprecipitated in wild type cells. NaSal decreased this interaction. Heatshock results in disruption of the interaction and the lack of HSP90 interaction. 4B, 4C, 4D, 4E, 4F, 5 S121 was phosphorylated in HSF1.