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/ Pyruvate kinase isozyme type M2 (PKM2) interacts and cooperates with Oct-4 in regulating transcription. Pyruvate kinase isozyme type M2 (PKM2) interacts and cooperates with Oct-4 in regulating transcription. Int. J. Biochem. Cell Biol. (1357-2725) 2007 Lee J., Kim HK., Han YM., Kim J. Cancer - Interactions investigated in the context of cancer Source of Oct-4 identified from PMID 1967980. jkim@sogang.ac.kr 01-April-2008: Contacted by JYOTI. davet 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 anti tag western anti tag western blot anti tag western Lee J., Kim HK., Han YM., Kim J. Cancer - Interactions investigated in the context of cancer Source of Oct-4 identified from PMID 1967980. jkim@sogang.ac.kr Accepted 2008-FEB-25 by DAVET. 01-April-2008: Contacted by JYOTI. Int. J. Biochem. Cell Biol. (1357-2725) 2007 davet Pyruvate kinase isozyme type M2 (PKM2) interacts and cooperates with Oct-4 in regulating transcription. Pyruvate kinase isozyme type M2 (PKM2) interacts and cooperates with Oct-4 in regulating transcription. Int. J. Biochem. Cell Biol. (1357-2725) 2007 Lee J., Kim HK., Han YM., Kim J. Cancer - Interactions investigated in the context of cancer Source of Oct-4 identified from PMID 1967980. jkim@sogang.ac.kr 01-April-2008: Contacted by JYOTI. davet in vitro In vitro pull down pull down ms participant Identification by mass spectrometry ms participant Lee J., Kim HK., Han YM., Kim J. Cancer - Interactions investigated in the context of cancer Source of Oct-4 identified from PMID 1967980. jkim@sogang.ac.kr Accepted 2008-FEB-25 by DAVET. 01-April-2008: Contacted by JYOTI. Int. J. Biochem. Cell Biol. (1357-2725) 2007 davet Pyruvate kinase isozyme type M2 (PKM2) interacts and cooperates with Oct-4 in regulating transcription. Pyruvate kinase isozyme type M2 (PKM2) interacts and cooperates with Oct-4 in regulating transcription. Int. J. Biochem. Cell Biol. (1357-2725) 2007 Lee J., Kim HK., Han YM., Kim J. Cancer - Interactions investigated in the context of cancer Source of Oct-4 identified from PMID 1967980. jkim@sogang.ac.kr 01-April-2008: Contacted by JYOTI. davet in vitro In vitro pull down pull down anti tag western anti tag western blot anti tag western Lee J., Kim HK., Han YM., Kim J. Cancer - Interactions investigated in the context of cancer Source of Oct-4 identified from PMID 1967980. jkim@sogang.ac.kr Accepted 2008-FEB-25 by DAVET. 01-April-2008: Contacted by JYOTI. Int. J. Biochem. Cell Biol. (1357-2725) 2007 davet Pyruvate kinase isozyme type M2 (PKM2) interacts and cooperates with Oct-4 in regulating transcription. Pyruvate kinase isozyme type M2 (PKM2) interacts and cooperates with Oct-4 in regulating transcription. Int. J. Biochem. Cell Biol. (1357-2725) 2007 Lee J., Kim HK., Han YM., Kim J. Cancer - Interactions investigated in the context of cancer Source of Oct-4 identified from PMID 1967980. jkim@sogang.ac.kr 01-April-2008: Contacted by JYOTI. davet chlae-cos_7 Cercopithecus aethiops simian cells transformed with SV40 Grivet cos_7 cercopithecus aethiops, by transformation with SV40 confocal microscopy confocal microscopy immunostaining immunostaining Lee J., Kim HK., Han YM., Kim J. Cancer - Interactions investigated in the context of cancer Source of Oct-4 identified from PMID 1967980. jkim@sogang.ac.kr Accepted 2008-FEB-25 by DAVET. 01-April-2008: Contacted by JYOTI. Int. J. Biochem. Cell Biol. (1357-2725) 2007 davet Pyruvate kinase isozyme type M2 (PKM2) interacts and cooperates with Oct-4 in regulating transcription. Pyruvate kinase isozyme type M2 (PKM2) interacts and cooperates with Oct-4 in regulating transcription. Int. J. Biochem. Cell Biol. (1357-2725) 2007 Lee J., Kim HK., Han YM., Kim J. Cancer - Interactions investigated in the context of cancer Source of Oct-4 identified from PMID 1967980. jkim@sogang.ac.kr 01-April-2008: Contacted by JYOTI. davet in vitro In vitro pull down pull down western blot western blot Immuno blot Immuno blot Lee J., Kim HK., Han YM., Kim J. Cancer - Interactions investigated in the context of cancer Source of Oct-4 identified from PMID 1967980. jkim@sogang.ac.kr Accepted 2008-FEB-25 by DAVET. 01-April-2008: Contacted by JYOTI. Int. J. Biochem. Cell Biol. (1357-2725) 2007 davet po5f1_mouse POU domain, class 5, transcription factor 1 Pou5f1 Oct-3 Oct-4 Otf-3 Otf3 Octamer-binding transcription factor 3 NF-A3 Octamer-binding protein 3 Octamer-binding protein 4 protein protein mouse Mus musculus Mouse MAGHLASDFAFSPPPGGGDGSAGLEPGWVDPRTWLSFQGPPGGPGIGPGSEVLGISPCPPAYEFCGGMAYCGPQVGLGLVPQVGVETLQPEGQAGARVESNSEGTSSEPCADRPNAVKLEKVEPTPEESQDMKALQKELEQFAKLLKQKRITLGYTQADVGLTLGVLFGKVFSQTTICRFEALQLSLKNMCKLRPLLEKWVEEADNNENLQEICKSETLVQARKRKRTSIENRVRWSLETMFLKCPKPSLQQITHIANQLGLEKDVVRVWFCNRRQKGKRSSIEYSQREEYEATGTPFPGGAVSFPLPPGPHFGTPGYGSPHFTTLYSVPFPEGEAFPSVPVTALGSPMHSN DIP protein P20263 original sequence version: 87 DIP protein P20263 original sequence version: 121 DIP protein P20263 original sequence version: 118 DIP protein P20263 original sequence version: 137 DIP protein P20263 original sequence version: 107 757E41DF52286714 kpym_mouse Pyruvate kinase PKM Pkm Pk3 Pkm2 Pykm Pyruvate kinase muscle isozyme protein protein mouse Mus musculus Mouse MPKPHSEAGTAFIQTQQLHAAMADTFLEHMCRLDIDSAPITARNTGIICTIGPASRSVEMLKEMIKSGMNVARLNFSHGTHEYHAETIKNVREATESFASDPILYRPVAVALDTKGPEIRTGLIKGSGTAEVELKKGATLKITLDNAYMEKCDENILWLDYKNICKVVEVGSKIYVDDGLISLQVKEKGADFLVTEVENGGSLGSKKGVNLPGAAVDLPAVSEKDIQDLKFGVEQDVDMVFASFIRKAADVHEVRKVLGEKGKNIKIISKIENHEGVRRFDEILEASDGIMVARGDLGIEIPAEKVFLAQKMMIGRCNRAGKPVICATQMLESMIKKPRPTRAEGSDVANAVLDGADCIMLSGETAKGDYPLEAVRMQHLIAREAEAAIYHLQLFEELRRLAPITSDPTEAAAVGAVEASFKCCSGAIIVLTKSGRSAHQVARYRPRAPIIAVTRNPQTARQAHLYRGIFPVLCKDAVLNAWAEDVDLRVNLAMDVGKARGFFKKGDVVIVLTGWRPGSGFTNTMRVVPVP DIP protein P52480 original sequence version: 159 34CBBC01BC0C047D pou5f1-pkm2 2 6 unspecified role unspecified role bait bait region sufficient to bind sufficient binding region sufficient to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain 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 ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role prey prey mouse-p19 Mus musculus embryonic teratocarcinoma cell line, epithelial-like, induced by retinoic acid p19 mouse embryonic teratocarcinoma cellline physical association physical association Fig. 1 we used affinity chromatography on immobilized Oct-4 (POU) together with MALDI-TOF (matrixassisted laser-desorption ionization-time-of-flight) MS (mass spectrometry) and isolated a novel Oct-4-interacting protein, pyruvate kinase type M2 (PKM2 or M2-PK). pkm2-pou5f1 5 6 unspecified role unspecified role prey prey mouse-p19 Mus musculus embryonic teratocarcinoma cell line, epithelial-like, induced by retinoic acid p19 mouse embryonic teratocarcinoma cellline 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 c-terminal 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 Fig. 2 A, 4 Association of Oct-4 with PKM2 in vitro. pou5f1-pkm2-1 1 7 unspecified role unspecified role prey prey region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 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 physical association physical association Fig. 2 C Co-affinity purification of Oct-4 with PKM2 from transfected cells. P52480; P20263 transfection pkm2-pou5f1-1 2 6 unspecified role unspecified role prey prey region his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined ecoli Escherichia coli (strain K12) 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 ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig. 2 D The interaction between Oct-4 and PKM2 is direct. pou5f1-pkm2-2 3 6 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 required to bind necessary binding region required to bind deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 7 unspecified role unspecified role prey prey region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined mouse-p19 Mus musculus embryonic teratocarcinoma cell line, epithelial-like, induced by retinoic acid p19 mouse embryonic teratocarcinoma cellline physical association physical association Fig. 3 PKM2 strongly bound to GSTOct-4 (POU) pkm2-pou5f1-2 4 6 unspecified role unspecified role neutral component neutral component 7 unspecified role unspecified role neutral component neutral component region flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined colocalization colocalization Fig. 5 A Overlay images indicated that Oct-4 and PKM2 in the nucleoplasm partially overlapped P20263; P52480 transfection