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/ B55?-associated PP2A complex controls PDK1-directed myc signaling and modulates rapamycin sensitivity in colorectal cancer. B55?-associated PP2A complex controls PDK1-directed myc signaling and modulates rapamycin sensitivity in colorectal cancer. Cancer Cell (1535-6108) 2010 imex curation Tan J., Lee PL., Li Z., Jiang X., Lim YC., Hooi SC., Yu Q. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer Chromatin - Epigenetic interactions resulting in chromatin modulation Fig 7B -Antibody to "PDK1" endogenous-(unphoph-ed variety) from BD Bioscineces: the SDS contains NO specificity info for the antibody, in spite of ~2 distinct bands visible on their QA piccys in WB. Authors confirm that PPP2RB coding region was obtained from Human colon tissue. Interactions containing PPP2A C have not been included as this molecule cannot be uniquely identified. yuq@gis.a-star.edu.sg 06-April-2011: Contacted by JYOTI. orchard Only protein-protein interactions human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Tan J., Lee PL., Li Z., Jiang X., Lim YC., Hooi SC., Yu Q. Apoptosis - Interactions involving proteins with a function related to apoptosis Fig 7B -Antibody to "PDK1" endogenous-(unphoph-ed variety) from BD Bioscineces: the SDS contains NO specificity info for the antibody, in spite of ~2 distinct bands visible on their QA piccys in WB. Authors confirm that PPP2RB coding region was obtained from Human colon tissue. Interactions containing PPP2A C have not been included as this molecule cannot be uniquely identified. yuq@gis.a-star.edu.sg Accepted 2011-JAN-24 by ORCHARD 06-April-2011: Contacted by JYOTI. Cancer Cell (1535-6108) 2010 orchard Only protein-protein interactions imex curation B55?-associated PP2A complex controls PDK1-directed myc signaling and modulates rapamycin sensitivity in colorectal cancer. B55?-associated PP2A complex controls PDK1-directed myc signaling and modulates rapamycin sensitivity in colorectal cancer. Cancer Cell (1535-6108) 2010 imex curation Tan J., Lee PL., Li Z., Jiang X., Lim YC., Hooi SC., Yu Q. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer Chromatin - Epigenetic interactions resulting in chromatin modulation Fig 7B -Antibody to "PDK1" endogenous-(unphoph-ed variety) from BD Bioscineces: the SDS contains NO specificity info for the antibody, in spite of ~2 distinct bands visible on their QA piccys in WB. Authors confirm that PPP2RB coding region was obtained from Human colon tissue. Interactions containing PPP2A C have not been included as this molecule cannot be uniquely identified. yuq@gis.a-star.edu.sg 06-April-2011: Contacted by JYOTI. orchard Only protein-protein interactions human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Tan J., Lee PL., Li Z., Jiang X., Lim YC., Hooi SC., Yu Q. Apoptosis - Interactions involving proteins with a function related to apoptosis Fig 7B -Antibody to "PDK1" endogenous-(unphoph-ed variety) from BD Bioscineces: the SDS contains NO specificity info for the antibody, in spite of ~2 distinct bands visible on their QA piccys in WB. HEK-TERV cells were used, which express both SV40 LT and mutant H-Ras-V12; they have constitutive activation of the MAPK-ERK pathway even in the absence of SV40 ST. Authors confirm that PPP2RB coding region was obtained from Human colon tissue. Interactions containing PPP2A C have not been included as this molecule cannot be uniquely identified. yuq@gis.a-star.edu.sg Accepted 2011-JAN-24 by ORCHARD 06-April-2011: Contacted by JYOTI. Cancer Cell (1535-6108) 2010 orchard Only protein-protein interactions imex curation B55?-associated PP2A complex controls PDK1-directed myc signaling and modulates rapamycin sensitivity in colorectal cancer. B55?-associated PP2A complex controls PDK1-directed myc signaling and modulates rapamycin sensitivity in colorectal cancer. Cancer Cell (1535-6108) 2010 imex curation Tan J., Lee PL., Li Z., Jiang X., Lim YC., Hooi SC., Yu Q. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer Chromatin - Epigenetic interactions resulting in chromatin modulation Fig 7B -Antibody to "PDK1" endogenous-(unphoph-ed variety) from BD Bioscineces: the SDS contains NO specificity info for the antibody, in spite of ~2 distinct bands visible on their QA piccys in WB. Authors confirm that PPP2RB coding region was obtained from Human colon tissue. Interactions containing PPP2A C have not been included as this molecule cannot be uniquely identified. yuq@gis.a-star.edu.sg 06-April-2011: Contacted by JYOTI. orchard Only protein-protein interactions human-293t Homo sapiens 293 cells transformed with SV40 large T antigen 293t 293 cells expressing SV40 large T antigen. The original designation of this cell line was 293tsA1609neo. It is not in Cabri,ATCC or HyperCLDB. anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Tan J., Lee PL., Li Z., Jiang X., Lim YC., Hooi SC., Yu Q. Apoptosis - Interactions involving proteins with a function related to apoptosis Fig 7B -Antibody to "PDK1" endogenous-(unphoph-ed variety) from BD Bioscineces: the SDS contains NO specificity info for the antibody, in spite of ~2 distinct bands visible on their QA piccys in WB. HEK-TERV cells were used, which express both SV40 LT and mutant H-Ras-V12; they have constitutive activation of the MAPK-ERK pathway even in the absence of SV40 ST. Authors confirm that PPP2RB coding region was obtained from Human colon tissue. Interactions containing PPP2A C have not been included as this molecule cannot be uniquely identified. yuq@gis.a-star.edu.sg Accepted 2011-JAN-24 by ORCHARD 06-April-2011: Contacted by JYOTI. Cancer Cell (1535-6108) 2010 orchard Only protein-protein interactions imex curation B55?-associated PP2A complex controls PDK1-directed myc signaling and modulates rapamycin sensitivity in colorectal cancer. B55?-associated PP2A complex controls PDK1-directed myc signaling and modulates rapamycin sensitivity in colorectal cancer. Cancer Cell (1535-6108) 2010 imex curation Tan J., Lee PL., Li Z., Jiang X., Lim YC., Hooi SC., Yu Q. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer Chromatin - Epigenetic interactions resulting in chromatin modulation Fig 7B -Antibody to "PDK1" endogenous-(unphoph-ed variety) from BD Bioscineces: the SDS contains NO specificity info for the antibody, in spite of ~2 distinct bands visible on their QA piccys in WB. Authors confirm that PPP2RB coding region was obtained from Human colon tissue. Interactions containing PPP2A C have not been included as this molecule cannot be uniquely identified. yuq@gis.a-star.edu.sg 06-April-2011: Contacted by JYOTI. orchard Only protein-protein interactions human-dld1 Human colorectal adenocarcinoma dld1 Human colon adenocarcinoma anti bait coip anti bait coimmunoprecipitation anti bait coip western blot western blot Immuno blot Immuno blot Tan J., Lee PL., Li Z., Jiang X., Lim YC., Hooi SC., Yu Q. Apoptosis - Interactions involving proteins with a function related to apoptosis Fig 7B -Antibody to "PDK1" endogenous-(unphoph-ed variety) from BD Bioscineces: the SDS contains NO specificity info for the antibody, in spite of ~2 distinct bands visible on their QA piccys in WB. Authors confirm that PPP2RB coding region was obtained from Human colon tissue. Interactions containing PPP2A C have not been included as this molecule cannot be uniquely identified. yuq@gis.a-star.edu.sg Accepted 2011-JAN-24 by ORCHARD 06-April-2011: Contacted by JYOTI. Cancer Cell (1535-6108) 2010 orchard Only protein-protein interactions imex curation B55?-associated PP2A complex controls PDK1-directed myc signaling and modulates rapamycin sensitivity in colorectal cancer. B55?-associated PP2A complex controls PDK1-directed myc signaling and modulates rapamycin sensitivity in colorectal cancer. Cancer Cell (1535-6108) 2010 imex curation Tan J., Lee PL., Li Z., Jiang X., Lim YC., Hooi SC., Yu Q. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer Chromatin - Epigenetic interactions resulting in chromatin modulation Fig 7B -Antibody to "PDK1" endogenous-(unphoph-ed variety) from BD Bioscineces: the SDS contains NO specificity info for the antibody, in spite of ~2 distinct bands visible on their QA piccys in WB. Authors confirm that PPP2RB coding region was obtained from Human colon tissue. Interactions containing PPP2A C have not been included as this molecule cannot be uniquely identified. yuq@gis.a-star.edu.sg 06-April-2011: Contacted by JYOTI. orchard Only protein-protein interactions human-dld1 Human colorectal adenocarcinoma dld1 Human colon adenocarcinoma anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Tan J., Lee PL., Li Z., Jiang X., Lim YC., Hooi SC., Yu Q. Apoptosis - Interactions involving proteins with a function related to apoptosis Fig 7B -Antibody to "PDK1" endogenous-(unphoph-ed variety) from BD Bioscineces: the SDS contains NO specificity info for the antibody, in spite of ~2 distinct bands visible on their QA piccys in WB. Fig 7B -Antibody to "PDK1" endogenous-(unphoph-ed variety) from BD Bioscineces: the SDS contains NO specificity info for the antibody, in spite of ~2 distinct bands visible on their QA piccys in WB. Fig 7B -Antibody to "PDK1" endogenous-(unphoph-ed variety) from BD Bioscineces: the SDS contains NO specificity info for the antibody, in spite of ~2 distinct bands visible on their QA piccys in WB. Fig 7B -Antibody to "PDK1" endogenous-(unphoph-ed variety) from BD Bioscineces: the SDS contains NO specificity info for the antibody, in spite of ~2 distinct bands visible on their QA piccys in WB. Authors confirm that PPP2RB coding region was obtained from Human colon tissue. Interactions containing PPP2A C have not been included as this molecule cannot be uniquely identified. yuq@gis.a-star.edu.sg Accepted 2011-JAN-24 by ORCHARD 06-April-2011: Contacted by JYOTI. Cancer Cell (1535-6108) 2010 orchard Only protein-protein interactions imex curation 2aaa_human Serine/threonine-protein phosphatase 2A 65 kDa regulatory subunit A alpha isoform PPP2R1A PP2A subunit A isoform PR65-alpha PP2A subunit A isoform R1-alpha Medium tumor antigen-associated 61 kDa protein protein protein human Homo sapiens Human MAAADGDDSLYPIAVLIDELRNEDVQLRLNSIKKLSTIALALGVERTRSELLPFLTDTIYDEDEVLLALAEQLGTFTTLVGGPEYVHCLLPPLESLATVEETVVRDKAVESLRAISHEHSPSDLEAHFVPLVKRLAGGDWFTSRTSACGLFSVCYPRVSSAVKAELRQYFRNLCSDDTPMVRRAAASKLGEFAKVLELDNVKSEIIPMFSNLASDEQDSVRLLAVEACVNIAQLLPQEDLEALVMPTLRQAAEDKSWRVRYMVADKFTELQKAVGPEITKTDLVPAFQNLMKDCEAEVRAAASHKVKEFCENLSADCRENVIMSQILPCIKELVSDANQHVKSALASVIMGLSPILGKDNTIEHLLPLFLAQLKDECPEVRLNIISNLDCVNEVIGIRQLSQSLLPAIVELAEDAKWRVRLAIIEYMPLLAGQLGVEFFDEKLNSLCMAWLVDHVYAIREAATSNLKKLVEKFGKEWAHATIIPKVLAMSGDPNYLHRMTTLFCINVLSEVCGQDITTKHMLPTVLRMAGDPVANVRFNVAKSLQKIGPILDNSTLQSEVKPILEKLTQDQDVDVKYFAQEALTVLSLA DIP protein P30153 original sequence version: 153 DIP protein P30153 original sequence version: 71 DIP protein P30153 original sequence version: 105 DIP protein P30153 original sequence version: 107 DIP protein P30153 original sequence version: 151 DIP protein P30153 original sequence version: 156 DIP protein P30153 original sequence version: 155 DIP protein P30153 original sequence version: 169 5174EBE94D537836 2abb_human Serine/threonine-protein phosphatase 2A 55 kDa regulatory subunit B beta isoform PP2A subunit B isoform beta PP2A subunit B isoform B55-beta PP2A subunit B isoform PR55-beta PPP2R2B PP2A subunit B isoform R2-beta protein protein human Homo sapiens Human MEEDIDTRKINNSFLRDHSYATEADIISTVEFNHTGELLATGDKGGRVVIFQREQESKNQVHRRGEYNVYSTFQSHEPEFDYLKSLEIEEKINKIRWLPQQNAAYFLLSTNDKTVKLWKVSERDKRPEGYNLKDEEGRLRDPATITTLRVPVLRPMDLMVEATPRRVFANAHTYHINSISVNSDYETYMSADDLRINLWNFEITNQSFNIVDIKPANMEELTEVITAAEFHPHHCNTFVYSSSKGTIRLCDMRASALCDRHTKFFEEPEDPSNRSFFSEIISSISDVKFSHSGRYIMTRDYLTVKVWDLNMENRPIETYQVHDYLRSKLCSLYENDCIFDKFECVWNGSDSVIMTGSYNNFFRMFDRNTKRDVTLEASRENSKPRAILKPRKVCVGGKRRKDEISVDSLDFSKKILHTAWHPSENIIAVAATNNLYIFQDKVN C383C834B2852B8F ks6b1_human Ribosomal protein S6 kinase beta-1 RPS6KB1 STK14A Ribosomal protein S6 kinase I p70 ribosomal S6 kinase alpha 70 kDa ribosomal protein S6 kinase 1 Serine/threonine-protein kinase 14A protein protein human Homo sapiens Human MRRRRRRDGFYPAPDFRDREAEDMAGVFDIDLDQPEDAGSEDELEEGGQLNESMDHGGVGPYELGMEHCEKFEISETSVNRGPEKIRPECFELLRVLGKGGYGKVFQVRKVTGANTGKIFAMKVLKKAMIVRNAKDTAHTKAERNILEEVKHPFIVDLIYAFQTGGKLYLILEYLSGGELFMQLEREGIFMEDTACFYLAEISMALGHLHQKGIIYRDLKPENIMLNHQGHVKLTDFGLCKESIHDGTVTHTFCGTIEYMAPEILMRSGHNRAVDWWSLGALMYDMLTGAPPFTGENRKKTIDKILKCKLNLPPYLTQEARDLLKKLLKRNAASRLGAGPGDAGEVQAHPFFRHINWEELLARKVEPPFKPLLQSEEDVSQFDSKFTRQTPVDSPDDSTLSESANQVFLGFTYVAPSVLESVKEKFSFEPKIRSPRRFIGSPRTPVSPVKFSPGDFWGRGASASTANPQTPVEYPMETSGIEQMDVTMSGEASAPLPIRQPNSGPYKKQAFPMISKRPEHLRMNL DIP protein P23443 original sequence version: 163 DIP protein P23443 original sequence version: 88 2C3BA13CCDAF4AB3 pdpk1_human 3-phosphoinositide-dependent protein kinase 1 PDPK1 PDK1 protein protein human Homo sapiens Human MARTTSQLYDAVPIQSSVVLCSCPSPSMVRTQTESSTPPGIPGGSRQGPAMDGTAAEPRPGAGSLQHAQPPPQPRKKRPEDFKFGKILGEGSFSTVVLARELATSREYAIKILEKRHIIKENKVPYVTRERDVMSRLDHPFFVKLYFTFQDDEKLYFGLSYAKNGELLKYIRKIGSFDETCTRFYTAEIVSALEYLHGKGIIHRDLKPENILLNEDMHIQITDFGTAKVLSPESKQARANSFVGTAQYVSPELLTEKSACKSSDLWALGCIIYQLVAGLPPFRAGNEYLIFQKIIKLEYDFPEKFFPKARDLVEKLLVLDATKRLGCEEMEGYGPLKAHPFFESVTWENLHQQTPPKLTAYLPAMSEDDEDCYGNYDNLLSQFGCMQVSSSSSSHSLSASDTGLPQRSGSNIEQYIHDLDSNSFELDLQFSEDEKRLLLEKQAGGNPWHQFVENNLILKMGPVDKRKGLFARRRQLLLTEGPHLYYVDPVNKVLKGEIPWSQELRPEAKNFKTFFVHTPNRTYYLMDPSGNAHKWCRKIQEVWRQRYQSHPDAAVQ DIP protein O15530 original sequence version: 169 DIP protein O15530-1 original sequence version: 178 ED8C0306DC4D0653 ppp2r2b_1a_c 5 7 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 unspecified role unspecified role prey prey physical association physical association Fig 3D doxycycline -a semi-synthetic tetracycline antibiotic. Subunit(s) of the catalytic subunit of PP2A were also present in the immunoprecipitation but could not be ascribed to a single gene product. pdpk1_ppp2r2b-1 5 9 unspecified role unspecified role bait bait 7 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 P67775 should be the paper's "PP2A C" - but may not be...PP2A A Subunit(81G5), -if this mol is PPP2R1A - then the ab detects 2 isoforms - so cannot use this in the interaction...(SDS states UniProt entry as P30153.)The PP2A C mol- cant see any antibody used to detect this, listed anywhere in the paper. PPP2R2B: searching the Upstate/now Millipore catalogue gives only ONE Ab -Catalogue Number : 07-334 - which gives the SwissP ID that I have already entered, deduced from the paper. However blasting the Immunogen sequence used by the supplier gives many different proteins - Q13362-3/2 amongst others....SDS states :Specificity: PP2A B' subunit, all isoforms (note the ALL isoforms - not just PPP2R2B...However the PPP2R2B molecule was transfected into the cells - so this must be the reacting entity- there is only one major band on the WB? (SO says this is OK!!- dont delete!)Is this still holding when PPP2R2B - myc is the PREY rather than the bait, as before?- both are bait and prey are co-transfected. Fig 7B. doxycycline -a semi-synthetic tetracycline antibiotic. ppp2r2b-pdk1 2 9 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait physical association physical association Fig 7C. The endogenous interaction between PDK1 and B55-beta was markedly reduced in HEKTERV cells expressing shPPP2R2B. s6k-ppp2r2b-pp2ac 5 6 unspecified role unspecified role prey prey 8 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait region myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined association association Fig S3B doxycycline -a semi-synthetic tetracycline antibiotic. s6k-2abb-pp2aa 5 8 unspecified role unspecified role bait bait 7 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 Fig S3A doxycycline -a semi-synthetic tetracycline antibiotic. The catalytic subunit(s) of PP2A were also present in the complex but could not be ascribed to a single gene product. ppp2r2b-pdpk1-1 1 9 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 7 unspecified role unspecified role bait bait 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 Fig 7A. doxycycline -a semi-synthetic tetracycline antibiotic. ppp2r2b-pdpk1 5 9 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait 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 Fig 7B. doxycycline -a semi-synthetic tetracycline antibiotic. pdpk1-ppp2r2b 1 7 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 9 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 physical association physical association Fig 7A. doxycycline -a semi-synthetic tetracycline antibiotic. pdpk1_ppp2r2b-6 4 9 unspecified role unspecified role bait bait 7 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 Fig 7B. doxycycline -a semi-synthetic tetracycline antibiotic. pdpk1_ppp2r2b-7 3 7 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait physical association physical association Fig 7C. The endogenous interaction between PDK1 and B55-beta was markedly reduced in HEKTERV cells expressing shPPP2R2B. pdpk1-ppp2r2b-1 2 7 unspecified role unspecified role prey prey 9 unspecified role unspecified role bait bait physical association physical association Fig 7C. The endogenous interaction between PDK1 and B55-beta was markedly reduced in HEKTERV cells expressing shPPP2R2B.