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/ Chaperoning checkpoint kinase 1 (Chk1), an Hsp90 client, with purified chaperones. Chaperoning checkpoint kinase 1 (Chk1), an Hsp90 client, with purified chaperones. J. Biol. Chem. (0021-9258) 2006 imex curation Arlander SJ., Felts SJ., Wagner JM., Stensgard B., Toft DO., Karnitz LM. BioCreative - Critical Assessment of Information Extraction systems in Biology Source of CDC25C not gven in paper and could not be traced but positions of key residues only match to the human sequence. cells were treated with 10mMhydroxyurea (HU) 1 h before lysis karnitz.larry@mayo.edu 13-SEP-2006: Contacted by JYOTI Only protein-protein interactions 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 bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Arlander SJ., Felts SJ., Wagner JM., Stensgard B., Toft DO., Karnitz LM. BioCreative - Critical Assessment of Information Extraction systems in Biology Source of CDC25C not gven in paper and could not be traced but positions of key residues only match to the human sequence. cells were treated with 10mMhydroxyurea (HU) 1 h before lysis karnitz.larry@mayo.edu Accepted 2006-AUG-1 by DAVET. 13-SEP-2006: Contacted by JYOTI J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation Chaperoning checkpoint kinase 1 (Chk1), an Hsp90 client, with purified chaperones. Chaperoning checkpoint kinase 1 (Chk1), an Hsp90 client, with purified chaperones. J. Biol. Chem. (0021-9258) 2006 imex curation Arlander SJ., Felts SJ., Wagner JM., Stensgard B., Toft DO., Karnitz LM. BioCreative - Critical Assessment of Information Extraction systems in Biology Source of CDC25C not gven in paper and could not be traced but positions of key residues only match to the human sequence. cells were treated with 10mMhydroxyurea (HU) 1 h before lysis karnitz.larry@mayo.edu 13-SEP-2006: Contacted by JYOTI Only protein-protein interactions in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Arlander SJ., Felts SJ., Wagner JM., Stensgard B., Toft DO., Karnitz LM. 1 �g of GST-Chk1-(1-265) was incubated with 2 �g of Hsp90, 20 �g of Hsp70, 4 �g of Ydj1, 4 �g of Cdc37, 5 �g of Hop, 5 �g of p23, and 0.14 units of CK2 at 30 �C for 90 min prior to pulldown. karnitz.larry@mayo.edu Accepted 2006-AUG-24 by DAVET. J. Biol. Chem. (0021-9258) 2006 Chaperoning checkpoint kinase 1 (Chk1), an Hsp90 client, with purified chaperones. Chaperoning checkpoint kinase 1 (Chk1), an Hsp90 client, with purified chaperones. J. Biol. Chem. (0021-9258) 2006 imex curation Arlander SJ., Felts SJ., Wagner JM., Stensgard B., Toft DO., Karnitz LM. BioCreative - Critical Assessment of Information Extraction systems in Biology Source of CDC25C not gven in paper and could not be traced but positions of key residues only match to the human sequence. cells were treated with 10mMhydroxyurea (HU) 1 h before lysis karnitz.larry@mayo.edu 13-SEP-2006: Contacted by JYOTI Only protein-protein interactions in vitro In vitro protein kinase assay protein kinase assay predetermined predetermined participant predetermined Arlander SJ., Felts SJ., Wagner JM., Stensgard B., Toft DO., Karnitz LM. BioCreative - Critical Assessment of Information Extraction systems in Biology Source of CDC25C not gven in paper and could not be traced but positions of key residues only match to the human sequence. cells were treated with 10mMhydroxyurea (HU) 1 h before lysis karnitz.larry@mayo.edu Accepted 2006-AUG-1 by DAVET. 13-SEP-2006: Contacted by JYOTI J. Biol. Chem. (0021-9258) 2006 Only protein-protein interactions imex curation hs90b_human Heat shock protein HSP 90-beta HSP90AB1 HSP90B HSPC2 HSPCB Heat shock 84 kDa protein protein human Homo sapiens Human MPEEVHHGEEEVETFAFQAEIAQLMSLIINTFYSNKEIFLRELISNASDALDKIRYESLTDPSKLDSGKELKIDIIPNPQERTLTLVDTGIGMTKADLINNLGTIAKSGTKAFMEALQAGADISMIGQFGVGFYSAYLVAEKVVVITKHNDDEQYAWESSAGGSFTVRADHGEPIGRGTKVILHLKEDQTEYLEERRVKEVVKKHSQFIGYPITLYLEKEREKEISDDEAEEEKGEKEEEDKDDEEKPKIEDVGSDEEDDSGKDKKKKTKKIKEKYIDQEELNKTKPIWTRNPDDITQEEYGEFYKSLTNDWEDHLAVKHFSVEGQLEFRALLFIPRRAPFDLFENKKKKNNIKLYVRRVFIMDSCDELIPEYLNFIRGVVDSEDLPLNISREMLQQSKILKVIRKNIVKKCLELFSELAEDKENYKKFYEAFSKNLKLGIHEDSTNRRRLSELLRYHTSQSGDEMTSLSEYVSRMKETQKSIYYITGESKEQVANSAFVERVRKRGFEVVYMTEPIDEYCVQQLKEFDGKSLVSVTKEGLELPEDEEEKKKMEESKAKFENLCKLMKEILDKKVEKVTISNRLVSSPCCIVTSTYGWTANMERIMKAQALRDNSTMGYMMAKKHLEINPDHPIVETLRQKAEADKNDKAVKDLVVLLFETALLSSGFSLEDPQTHSNRIYRMIKLGLGIDEDEVAAEEPNAAVPDEIPPLEGDEDASRMEEVD DIP protein P08238 original sequence version: 91 DIP protein P08238 original sequence version: 149 DIP protein P08238 original sequence version: 165 DIP protein P08238 original sequence version: 167 DIP protein P08238 original sequence version: 210 DIP protein P08238 original sequence version: 187 A93118C214D03810 mpip3_human M-phase inducer phosphatase 3 CDC25C Dual specificity phosphatase Cdc25C protein protein human Homo sapiens Human MSTELFSSTREEGSSGSGPSFRSNQRKMLNLLLERDTSFTVCPDVPRTPVGKFLGDSANLSILSGGTPKRCLDLSNLSSGEITATQLTTSADLDETGHLDSSGLQEVHLAGMNHDQHLMKCSPAQLLCSTPNGLDRGHRKRDAMCSSSANKENDNGNLVDSEMKYLGSPITTVPKLDKNPNLGEDQAEEISDELMEFSLKDQEAKVSRSGLYRSPSMPENLNRPRLKQVEKFKDNTIPDKVKKKYFSGQGKLRKGLCLKKTVSLCDITITQMLEEDSNQGHLIGDFSKVCALPTVSGKHQDLKYVNPETVAALLSGKFQGLIEKFYVIDCRYPYEYLGGHIQGALNLYSQEELFNFFLKKPIVPLDTQKRIIIVFHCEFSSERGPRMCRCLREEDRSLNQYPALYYPELYILKGGYRDFFPEYMELCEPQSYCPMHHQDHKTELLRCRSQSKVQEGERQLREQIALLVKDMSP DIP protein P30307 original sequence version: 148 DIP protein P30307 original sequence version: 88 DIP protein P30307 original sequence version: 112 DIP protein P30307 original sequence version: 108 0658A1F1B9B8996A raf1_human RAF proto-oncogene serine/threonine-protein kinase RAF1 RAF Proto-oncogene c-RAF Raf-1 protein protein human Homo sapiens Human MEHIQGAWKTISNGFGFKDAVFDGSSCISPTIVQQFGYQRRASDDGKLTDPSKTSNTIRVFLPNKQRTVVNVRNGMSLHDCLMKALKVRGLQPECCAVFRLLHEHKGKKARLDWNTDAASLIGEELQVDFLDHVPLTTHNFARKTFLKLAFCDICQKFLLNGFRCQTCGYKFHEHCSTKVPTMCVDWSNIRQLLLFPNSTIGDSGVPALPSLTMRRMRESVSRMPVSSQHRYSTPHAFTFNTSSPSSEGSLSQRQRSTSTPNVHMVSTTLPVDSRMIEDAIRSHSESASPSALSSSPNNLSPTGWSQPKTPVPAQRERAPVSGTQEKNKIRPRGQRDSSYYWEIEASEVMLSTRIGSGSFGTVYKGKWHGDVAVKILKVVDPTPEQFQAFRNEVAVLRKTRHVNILLFMGYMTKDNLAIVTQWCEGSSLYKHLHVQETKFQMFQLIDIARQTAQGMDYLHAKNIIHRDMKSNNIFLHEGLTVKIGDFGLATVKSRWSGSQQVEQPTGSVLWMAPEVIRMQDNNPFSFQSDVYSYGIVLYELMTGELPYSHINNRDQIIFMVGRGYASPDLSKLYKNCPKAMKRLVADCVKKVKEERPLFPQILSSIELLQHSLPKINRSASEPSLHRAAHTEDINACTLTTSPRLPVF DIP protein P04049 original sequence version: 117 DIP protein P04049 original sequence version: 106 DIP protein P04049 original sequence version: 114 DIP protein P04049 original sequence version: 141 DIP protein P04049 original sequence version: 142 DIP protein P04049 original sequence version: 147 DIP protein P04049-1 original sequence version: 136 DIP protein P04049 original sequence version: 136 DIP protein P04049 original sequence version: 153 DIP protein P04049 original sequence version: 150 DIP protein P04049 original sequence version: 164 DIP protein P04049 original sequence version: 172 DIP protein P04049-1 original sequence version: 172 DIP protein P04049 original sequence version: 184 DIP protein P04049 original sequence version: 188 DIP protein P04049 original sequence version: 195 EF821B5349711BC3 chk1_human Serine/threonine-protein kinase Chk1 CHEK1 CHK1 CHK1 checkpoint homolog Checkpoint kinase-1 Cell cycle checkpoint kinase protein protein human Homo sapiens Human MAVPFVEDWDLVQTLGEGAYGEVQLAVNRVTEEAVAVKIVDMKRAVDCPENIKKEICINKMLNHENVVKFYGHRREGNIQYLFLEYCSGGELFDRIEPDIGMPEPDAQRFFHQLMAGVVYLHGIGITHRDIKPENLLLDERDNLKISDFGLATVFRYNNRERLLNKMCGTLPYVAPELLKRREFHAEPVDVWSCGIVLTAMLAGELPWDQPSDSCQEYSDWKEKKTYLNPWKKIDSAPLALLHKILVENPSARITIPDIKKDRWYNKPLKKGAKRPRVTSGGVSESPSGFSKHIQSNLDFSPVNSASSEENVKYSSSQPEPRTGLSLWDTSPSYIDKLVQGISFSQPTCPDHMLLNSQLLGTPGSSQNPWQRLVKRMTRFFTKLDADKSYQCLKETCEKLGYQWKKSCMNQVTISTTDRRNNKLIFKVNLLEMDDKILVDFRLSKGDGLEFKRHFLKIKGKLIDIVSSQKIWLPAT DIP protein O14757 original sequence version: 102 0ABD0FAB67E60F67 hspcb-raf1-1 1 4 unspecified role unspecified role bait bait 6 unspecified role unspecified role prey prey physical association physical association We first demonstrated that covalent coupling did not disrupt the ability of Hsp90 to co-precipitate Raf-1, a bona fide Hsp90 client 1 cdc25c-chek1-1 3 7 enzyme enzyme unspecified role unspecified role region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined in vitro In vitro 5 enzyme target enzyme target substrate substrate unspecified role unspecified role 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 deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation Phosphorylation of CDC25C In vitro transcription/translation reactions were programmed with empty vector or a vector that expresses HA-tagged full-length Chk1 and immunoprecipitated with anti-HA antibody conjugated to agarose beads. Following washing, the precipitates were subjected to kinase assays by incubating with [-32P]ATP and the GST-Cdc25C substrate. 2A hspcb-chk1-1 2 4 unspecified role unspecified role prey prey 7 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 deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) physical association physical association Active Chk1 is associated with Hsp90. 7