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/ hu-2007-1 Efficient p53 activation and apoptosis by simultaneous disruption of binding to MDM2 and MDMX. in vitro In vitro phage display phage display predetermined predetermined participant predetermined Hu B., Gilkes DM., Chen J. Apoptosis - Interactions involving proteins with a function related to apoptosis Jiandong.chen@moffitt.org Cancer Res. (0008-5472) 2007 Cancer - Interactions investigated in the context of cancer hu-2007-4 Efficient p53 activation and apoptosis by simultaneous disruption of binding to MDM2 and MDMX. human-mcf7 Homo sapiens mammary carcinoma cells mcf7 mammary carcinoma cells anti tag coip anti tag coimmunoprecipitation anti tag coip western blot western blot Immuno blot Immuno blot Hu B., Gilkes DM., Chen J. Apoptosis - Interactions involving proteins with a function related to apoptosis Jiandong.chen@moffitt.org Cancer Res. (0008-5472) 2007 Cancer - Interactions investigated in the context of cancer hu-2007-2 Efficient p53 activation and apoptosis by simultaneous disruption of binding to MDM2 and MDMX. in vitro In vitro elisa enzyme linked immunosorbent assay ELISA ELISA elisa antibody detection identification by antibody antibody detection Hu B., Gilkes DM., Chen J. Apoptosis - Interactions involving proteins with a function related to apoptosis Jiandong.chen@moffitt.org Cancer Res. (0008-5472) 2007 Cancer - Interactions investigated in the context of cancer hu-2007-3 Efficient p53 activation and apoptosis by simultaneous disruption of binding to MDM2 and MDMX. in vitro In vitro pull down pull down weight autoradiogra molecular weight estimation by autoradiography weight autoradiogra Hu B., Gilkes DM., Chen J. Apoptosis - Interactions involving proteins with a function related to apoptosis Jiandong.chen@moffitt.org Cancer Res. (0008-5472) 2007 Cancer - Interactions investigated in the context of cancer mdm4_human Protein Mdm4 MDM4 MDMX p53-binding protein Mdm4 Mdm2-like p53-binding protein Protein Mdmx Double minute 4 protein protein protein human Homo sapiens Human MTSFSTSAQCSTSDSACRISPGQINQVRPKLPLLKILHAAGAQGEMFTVKEVMHYLGQYIMVKQLYDQQEQHMVYCGGDLLGELLGRQSFSVKDPSPLYDMLRKNLVTLATATTDAAQTLALAQDHSMDIPSQDQLKQSAEESSTSRKRTTEDDIPTLPTSEHKCIHSREDEDLIENLAQDETSRLDLGFEEWDVAGLPWWFLGNLRSNYTPRSNGSTDLQTNQDVGTAIVSDTTDDLWFLNESVSEQLGVGIKVEAADTEQTSEEVGKVSDKKVIEVGKNDDLEDSKSLSDDTDVEVTSEDEWQCTECKKFNSPSKRYCFRCWALRKDWYSDCSKLTHSLSTSDITAIPEKENEGNDVPDCRRTISAPVVRPKDAYIKKENSKLFDPCNSVEFLDLAHSSESQETISSMGEQLDNLSEQRTDTENMEDCQNLLKPCSLCEKRPRDGNIIHGRTGHLVTCFHCARRLKKAGASCPICKKEIQLVIKVFIA t_mx_1_random_pept T-MX-1 a random 12-mer peptide peptide peptide polypeptide oligopeptide oligopeptide polypeptide chemical synthesis Chemical synthesis (Chemical synthesis) FAPLNRTVETSP An amino acid sequence which is not represented in UniProt A random 12-mer peptide used in a phage display assay. mdm2_human E3 ubiquitin-protein ligase Mdm2 MDM2 p53-binding protein Mdm2 Oncoprotein Mdm2 Double minute 2 protein RING-type E3 ubiquitin transferase Mdm2 protein protein human Homo sapiens Human MCNTNMSVPTDGAVTTSQIPASEQETLVRPKPLLLKLLKSVGAQKDTYTMKEVLFYLGQYIMTKRLYDEKQQHIVYCSNDLLGDLFGVPSFSVKEHRKIYTMIYRNLVVVNQQESSDSGTSVSENRCHLEGGSDQKDLVQELQEEKPSSSHLVSRPSTSSRRRAISETEENSDELSGERQRKRHKSDSISLSFDESLALCVIREICCERSSSSESTGTPSNPDLDAGVSEHSGDWLDQDSVSDQFSVEFEVESLDSEDYSLSEEGQELSDEDDEVYQVTVYQAGESDTDSFEEDPEISLADYWKCTSCNEMNPPLPSHCNRCWALRENWLPEDKGKDKGEISEKAKLENSTQAEEGFDVPDCKKTIVNDSRESCVEENDDKITQASQSQESEDYSQPSTSSSIIYSSQEDVKEFEREETQDKEESVESSLPLNAIEPCVICQGRPKNGCIVHGKTGHLMACFTCAKKLKKRNKPCPVCRQPIQMIVLTYFP pdi_random_peptide pDI peptide (random 12-mer) peptide peptide polypeptide oligopeptide oligopeptide polypeptide chemical synthesis Chemical synthesis (Chemical synthesis) LTFEHYWAQLTS An amino acid sequence which is not represented in UniProt A random 12-mer used in phage display assay. t_mx_4_random_pept T-MX-4 a random 12-mer peptide peptide peptide polypeptide oligopeptide oligopeptide polypeptide chemical synthesis Chemical synthesis (Chemical synthesis) YAVSSSPRVAAL An amino acid sequence which is not represented in UniProt A random 12-mer peptide used in a phage display assay. t_mx_9_random_pept T-MX-9 a random 12-mer peptide peptide peptide polypeptide oligopeptide oligopeptide polypeptide chemical synthesis Chemical synthesis (Chemical synthesis) VVHVPNSATPPR An amino acid sequence which is not represented in UniProt A random 12-mer peptide used in a phage display assay. t_m2_1_random_pept T-M2-1 a random 12-mer peptide peptide peptide polypeptide oligopeptide oligopeptide polypeptide chemical synthesis Chemical synthesis (Chemical synthesis) QQMHLMSYAPGP An amino acid sequence which is not represented in UniProt t_m2_2_random_pept T-M2-2 a random 12-mer peptide peptide peptide polypeptide oligopeptide oligopeptide polypeptide chemical synthesis Chemical synthesis (Chemical synthesis) TIRPSTTMDSPT An amino acid sequence which is not represented in UniProt A random 12-mer peptide used in a phage display assay. t_m2_3_random_pept T-M2-3 random 12-mer peptide peptide peptide polypeptide oligopeptide oligopeptide polypeptide chemical synthesis Chemical synthesis (Chemical synthesis) YANPQMEKAFES An amino acid sequence which is not represented in UniProt A random 12-mer peptide used in a phage display assay. t_m2_5_random_pept T-M2-5 random 12-mer peptide peptide peptide polypeptide oligopeptide oligopeptide polypeptide chemical synthesis Chemical synthesis (Chemical synthesis) LPNLTWALMPGA An amino acid sequence which is not represented in UniProt A random 12-mer peptide used in a phage display assay. t_m2_6_random_pept T-MX-6 random 12-mer peptide peptide peptide polypeptide oligopeptide oligopeptide polypeptide chemical synthesis Chemical synthesis (Chemical synthesis) YANPQMEKAFAS An amino acid sequence which is not represented in UniProt A random 12-mer peptide used in a phage display assay. t_m2_9_random_pept T-M2-9 random 12-mer peptide peptide peptide polypeptide oligopeptide oligopeptide polypeptide chemical synthesis Chemical synthesis (Chemical synthesis) LLADTTHHRPWT An amino acid sequence which is not represented in UniProt A random 12-mer peptide used in a phage display assay. thioredoxin_pdi_ecol E. coli Thioredoxin pDI fusion protein protein protein ecolx Escherichia coli MSDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGLTFEHYWAQLTSPCKMIAPILDEIADEYQGKLTVAKLNIDQNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQLKEFLDANLA An amino acid sequence which is not represented in UniProt The authors state that the pDI sequence is inserted between Gly33 and Pro34 but in the UniProtKB entry for E.coli thioredoxin Gly is at position 34 and Pro at 35. p53_human Cellular tumor antigen p53 TP53 P53 Tumor suppressor p53 Phosphoprotein p53 Antigen NY-CO-13 protein protein human Homo sapiens Human MEEPQSDPSVEPPLSQETFSDLWKLLPENNVLSPLPSQAMDDLMLSPDDIEQWFTEDPGPDEAPRMPEAAPPVAPAPAAPTPAAPAPAPSWPLSSSVPSQKTYQGSYGFRLGFLHSGTAKSVTCTYSPALNKMFCQLAKTCPVQLWVDSTPPPGTRVRAMAIYKQSQHMTEVVRRCPHHERCSDSDGLAPPQHLIRVEGNLRVEYLDDRNTFRHSVVVPYEPPEVGSDCTTIHYNYMCNSSCMGGMNRRPILTIITLEDSSGNLLGRNSFEVRVCACPGRDRRTEEENLRKKGEPHHELPPGSTKRALPNNTSSSPQPKKKPLDGEYFTLQIRGRERFEMFRELNEALELKDAQAGKEPGGSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSD mdm4-t_mx_1_random GST-MDM4 interacts with random 12-mer peptide T-MX-1 in a phage display 669799 N/A 669803 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 n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 669804 unspecified role unspecified role prey prey physical association physical association false false false Table 1 mdm2-pdi_random_pept GST-MDM2 interacts with random 12-mer peptide pDI in a phage display 669799 N/A 669805 unspecified role unspecified role bait bait n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 ecoli Escherichia coli (strain K12) N/A 669806 unspecified role unspecified role prey prey physical association physical association false false false Table 1 mdm4-t_mx_4_random GST-MDM4 interacts with random 12-mer peptide T-MX-4 in a phage display 669799 N/A 669807 unspecified role unspecified role prey prey N/A 669803 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 n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) physical association physical association false false false Table 1 mdm4-t_mx_9_random GST-MDM4 interacts with random 12-mer peptide T-MX-9 in a phage display 669799 N/A 669803 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 n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 669808 unspecified role unspecified role prey prey physical association physical association false false false Table 1 mdm4-pdi_random_pept GST-MDM4 interacts with random 12-mer peptide pDI in a phage display 669799 N/A 669803 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 n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 669806 unspecified role unspecified role prey prey physical association physical association false false false Table 1 mdm2-t_m2_1_random GST-MDM2 interacts with random 12-mer peptide T-M2-1 in a phage display 669799 N/A 669805 unspecified role unspecified role bait bait n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 ecoli Escherichia coli (strain K12) N/A 669809 unspecified role unspecified role prey prey physical association physical association false false false Table 1 mdm2-t_m2_2_random GST-MDM2 interacts with random 12-mer peptide T-M2-2 in a phage display 669799 N/A 669805 unspecified role unspecified role bait bait n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 ecoli Escherichia coli (strain K12) N/A 669810 unspecified role unspecified role prey prey physical association physical association false false false Table 1 mdm2-t_m2_3_random-2 GST-MDM2 interacts with random 12-mer peptide T-M2-3 in a phage display 669799 N/A 669805 unspecified role unspecified role bait bait n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 ecoli Escherichia coli (strain K12) N/A 669811 unspecified role unspecified role prey prey physical association physical association false false false Table 1 mdm2-t_m2_5_random-3 GST-MDM2 interacts with random 12-mer peptide T-M2-5 in a phage display 669799 N/A 669812 unspecified role unspecified role prey prey N/A 669805 unspecified role unspecified role bait bait n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 ecoli Escherichia coli (strain K12) physical association physical association false false false Table 1 mdm2-t_m2_6_random-4 GST-MDM2 interacts with random 12-mer peptide T-M2-6 in a phage display 669799 N/A 669805 unspecified role unspecified role bait bait n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 ecoli Escherichia coli (strain K12) N/A 669813 unspecified role unspecified role prey prey physical association physical association false false false Table 1 mdm2-t_m2_9_random-5 GST-MDM2 interacts with random 12-mer peptide T-M2-9 in a phage display 669799 N/A 669805 unspecified role unspecified role bait bait n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false 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 ecoli Escherichia coli (strain K12) N/A 669814 unspecified role unspecified role prey prey physical association physical association false false false Table 1 thioredoxin_pdi-mdm2 FLAG-DI coimmunoprecipitates endogenous Mdm2 in MCF-7 lysates 669800 N/A 669805 unspecified role unspecified role prey prey N/A 669815 unspecified role unspecified role bait bait phe37ala,trp41ala,leu44ala mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false n-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range false physical association physical association false false false 2 C P0AA25 infection thioredoxin_pdi-mdm4 FLAG-DI coimmunoprecipitates endogenous Mdm4 in MCF-7 lysates 669800 N/A 669815 unspecified role unspecified role bait bait phe37ala,trp41ala,leu44ala mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false certain certain sequence position certain certain certain sequence position certain false n-terminal flag tag flag tag FLAG-tagged FLAG DYKDDDDKV epitope tag DYKDDDDKV epitope tag FLAG FLAG-tagged n-term range n-terminal range n-term range n-term range n-terminal range n-term range false N/A 669803 unspecified role unspecified role prey prey physical association physical association false false false 2 C P0AA25 infection tp53-mdm2-2 p53 coimmunoprecipitates Mdm2 from MCF-7 lysates 669800 N/A 669803 unspecified role unspecified role prey prey N/A 669805 unspecified role unspecified role prey prey N/A 669816 unspecified role unspecified role bait bait association association false false false 2 D FLAG-DI - This interaction is inhibited by the E. coli thioredoxin-pDI chimeric protein, FLAG-DI but not FLAG-3A (in which three hydrophobic residues of the pDI sequence are mutated to alanine). Nutlin - This interaction is inhibited by Nutlin. tp53-mdm2 pDI random peptide inhibits the interaction between MDM2 and p53 as shown in an ELISA assay 669801 N/A 669816 unspecified role unspecified role bait bait 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 false ecoli Escherichia coli (strain K12) N/A 669805 unspecified role unspecified role prey prey 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 ecoli Escherichia coli (strain K12) physical association physical association false false false 1 A pDI random peptide - This interaction was inhibited with IC50 of 10 nmol/L by pDI random 12-mer peptide (LTFEHYWAQLTS). Mutation of three key hydrophobic residues in pDI to alanine abrogated the inhibition of complex formation by pDI, suggesting pDI mimics p53 binding to MDM2. p53pep - This interaction was inhibited with IC50 3.0 micromol/L by p53pep (QETFSDLWKLLP). 12/1 - This interaction was inhibited with IC50 0.15 micromol/L by 12/1 (MPRFMDYWEGLN). Nutlin - This interaction was inhibited with IC50 0.60 micromol/L by Nutlin. tp53-mdm4 pDI random peptide inhibits the interaction between MDMX and p53 as shown in an ELISA assay 669801 N/A 669803 unspecified role unspecified role prey prey 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 ecoli Escherichia coli (strain K12) N/A 669816 unspecified role unspecified role bait bait 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 false ecoli Escherichia coli (strain K12) physical association physical association false false false 1 A pDI random peptide - This interaction was inhibited with IC50 of 100 nmol/L by pDI random 12-mer peptide (LTFEHYWAQLTS). Mutation of three key hydrophobic residues in pDI to alanine abrogated the inhibition of complex formation by pDI, suggesting pDI mimics p53 binding to MDMX. p53pep - This interaction was inhibited with IC50 27.50 micromol/L by p53pep (QETFSDLWKLLP). 12/1 - This interaction was inhibited with IC50 1.25 micromol/L by 12/1 (MPRFMDYWEGLN). This interaction was not inhibited by Nutlin. tp53-mdm2-1 pDI peptide inhibits GST-p53(1-52) pull down of MDM2 669802 N/A 669816 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 n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) N/A 669805 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false in vitro In vitro physical association physical association false false false 1 C, D pDI random peptide - This interaction was inhibited with IC50 by pDI random 12-mer peptide (LTFEHYWAQLTS). Nutlin - This interaction was inhibited by Nutlin. Inhibition increased with concentration of Nutlin. tp53-mdm4-1 pDI peptide inhibits GST-p53(1-52) pull down of MDMX 669802 N/A 669803 unspecified role unspecified role prey prey region 35s radiolabel 35s radiolabel 35S s35 radiolabelled S35 s35 radiolabelled 35S S35 undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined false in vitro In vitro N/A 669816 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 n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain false ecoli Escherichia coli (strain K12) physical association physical association false false false 1 C, D pDI random peptide - This interaction was inhibited with IC50 by pDI random 12-mer peptide (LTFEHYWAQLTS). Nutlin did not inhibit this interaction.