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/ Efficient p53 activation and apoptosis by simultaneous disruption of binding to MDM2 and MDMX. Efficient p53 activation and apoptosis by simultaneous disruption of binding to MDM2 and MDMX. Cancer Res. (0008-5472) 2007 Hu B., Gilkes DM., Chen J. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer Jiandong.chen@moffitt.org 13-Nov-2007: Contacted by JYOTI. davet 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 Accepted 2007-NOV-18 by DAVET. Cancer Res. (0008-5472) 2007 davet Efficient p53 activation and apoptosis by simultaneous disruption of binding to MDM2 and MDMX. Efficient p53 activation and apoptosis by simultaneous disruption of binding to MDM2 and MDMX. Cancer Res. (0008-5472) 2007 Hu B., Gilkes DM., Chen J. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer Jiandong.chen@moffitt.org 13-Nov-2007: Contacted by JYOTI. davet 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 Accepted 2007-NOV-18 by DAVET. 13-Nov-2007: Contacted by JYOTI. Cancer Res. (0008-5472) 2007 davet Efficient p53 activation and apoptosis by simultaneous disruption of binding to MDM2 and MDMX. Efficient p53 activation and apoptosis by simultaneous disruption of binding to MDM2 and MDMX. Cancer Res. (0008-5472) 2007 Hu B., Gilkes DM., Chen J. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer Jiandong.chen@moffitt.org 13-Nov-2007: Contacted by JYOTI. davet 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 Accepted 2007-NOV-18 by DAVET. 13-Nov-2007: Contacted by JYOTI. Cancer Res. (0008-5472) 2007 davet Efficient p53 activation and apoptosis by simultaneous disruption of binding to MDM2 and MDMX. Efficient p53 activation and apoptosis by simultaneous disruption of binding to MDM2 and MDMX. Cancer Res. (0008-5472) 2007 Hu B., Gilkes DM., Chen J. Apoptosis - Interactions involving proteins with a function related to apoptosis Cancer - Interactions investigated in the context of cancer Jiandong.chen@moffitt.org 13-Nov-2007: Contacted by JYOTI. davet 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 Accepted 2007-NOV-18 by DAVET. 13-Nov-2007: Contacted by JYOTI. Cancer Res. (0008-5472) 2007 davet 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. 9BE0B5D7A281B1B5 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. 36C25DBD23C5B457 p53_human Cellular tumor antigen p53 TP53 P53 Tumor suppressor p53 Phosphoprotein p53 Antigen NY-CO-13 protein protein human Homo sapiens Human MEEPQSDPSVEPPLSQETFSDLWKLLPENNVLSPLPSQAMDDLMLSPDDIEQWFTEDPGPDEAPRMPEAAPPVAPAPAAPTPAAPAPAPSWPLSSSVPSQKTYQGSYGFRLGFLHSGTAKSVTCTYSPALNKMFCQLAKTCPVQLWVDSTPPPGTRVRAMAIYKQSQHMTEVVRRCPHHERCSDSDGLAPPQHLIRVEGNLRVEYLDDRNTFRHSVVVPYEPPEVGSDCTTIHYNYMCNSSCMGGMNRRPILTIITLEDSSGNLLGRNSFEVRVCACPGRDRRTEEENLRKKGEPHHELPPGSTKRALPNNTSSSPQPKKKPLDGEYFTLQIRGRERFEMFRELNEALELKDAQAGKEPGGSRAHSSHLKSKKGQSTSRHKKLMFKTEGPDSD DIP protein P04637 original sequence version: 121 DIP protein P04637 original sequence version: 115 DIP protein P04637-1 original sequence version: 224 DIP protein P04637 original sequence version: 117 DIP protein P04637-1 original sequence version: 226 DIP protein P04637 original sequence version: 226 DIP protein P04637 original sequence version: 212 DIP protein P04637 original sequence version: 220 DIP protein P04637-1 original sequence version: 220 DIP protein P04637-1 original sequence version: 145 DIP protein P04637 original sequence version: 146 DIP protein P04637 original sequence version: 145 DIP protein P04637 original sequence version: 166 DIP protein P04637-1 original sequence version: 166 DIP protein P04637 original sequence version: 169 DIP protein P04637 original sequence version: 177 DIP protein P04637 original sequence version: 175 DIP protein P04637-1 original sequence version: 175 DIP protein P04637-1 original sequence version: 172 DIP protein P04637-1 original sequence version: 169 DIP protein P04637 original sequence version: 170 DIP protein P04637 original sequence version: 172 DIP protein P04637-1 original sequence version: 194 DIP protein P04637 original sequence version: 196 DIP protein P04637 original sequence version: 185 DIP protein P04637-1 original sequence version: 191 DIP protein P04637 original sequence version: 191 DIP protein P04637 original sequence version: 180 DIP protein P04637-1 original sequence version: 197 DIP protein P04637 original sequence version: 183 DIP protein P04637-1 original sequence version: 183 DIP protein P04637-1 original sequence version: 185 DIP protein P04637 original sequence version: 199 DIP protein P04637 original sequence version: 201 DIP protein P04637 original sequence version: 189 DIP protein P04637 original sequence version: 235 DIP protein P04637 original sequence version: 230 DIP protein P04637 original sequence version: 234 DIP protein P04637-1 original sequence version: 234 DIP protein P04637-1 original sequence version: 235 DIP protein P04637 original sequence version: 239 DIP protein P04637-1 original sequence version: 212 DIP protein P04637-1 original sequence version: 213 DIP protein P04637 original sequence version: 213 DIP protein P04637 original sequence version: 206 DIP protein P04637-1 original sequence version: 188 DIP protein P04637-1 original sequence version: 228 DIP protein P04637 original sequence version: 208 DIP protein P04637 original sequence version: 240 DIP protein P04637-1 original sequence version: 240 DIP protein P04637 original sequence version: 204 DIP protein P04637-1 original sequence version: 204 DIP protein P04637-1 original sequence version: 209 DIP protein P04637-1 original sequence version: 203 DIP protein P04637 original sequence version: 221 DIP protein P04637-1 original sequence version: 152 DIP protein P04637 original sequence version: 217 DIP protein P04637-1 original sequence version: 140 DIP protein P04637 original sequence version: 215 DIP protein P04637 original sequence version: 214 DIP protein P04637-1 original sequence version: 182 DIP protein P04637 original sequence version: 182 DIP protein P04637 original sequence version: 188 DIP protein P04637 original sequence version: 192 DIP protein P04637-1 original sequence version: 192 DIP protein P04637 original sequence version: 218 DIP protein P04637-1 original sequence version: 218 AD5C149FD8106131 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. C5F17F86AE3B1686 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. F8E6396CA1C36B51 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. 8EB00389A91775A5 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 405F1A7C9E45B697 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. 87F8D428EB2DC361 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. FF677788301935D0 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 DIP protein O15151 original sequence version: 141 DIP protein O15151 original sequence version: 109 DIP protein O15151 original sequence version: 129 DIP protein O15151 original sequence version: 147 415E6FA5A0C69857 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 DIP protein Q00987 original sequence version: 93 DIP protein Q00987-1 original sequence version: 192 DIP protein Q00987 original sequence version: 88 DIP protein Q00987 original sequence version: 180 DIP protein Q00987 original sequence version: 188 DIP protein Q00987 original sequence version: 189 DIP protein Q00987 original sequence version: 115 DIP protein Q00987-1 original sequence version: null DIP protein Q00987 original sequence version: 149 DIP protein Q00987 original sequence version: 146 DIP protein Q00987 original sequence version: 140 DIP protein Q00987 original sequence version: 183 DIP protein Q00987 original sequence version: 145 DIP protein Q00987 original sequence version: 182 DIP protein Q00987 original sequence version: 151 DIP protein Q00987 original sequence version: 156 DIP protein Q00987-1 original sequence version: 160 DIP protein Q00987 original sequence version: 160 DIP protein Q00987 original sequence version: 168 DIP protein Q00987 original sequence version: 186 DIP protein Q00987 original sequence version: 206 F37CE163876BC983 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. C5F7BF86AE3B1686 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. 91B73C567DE1F1B1 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. 1F4CE1F8DCD1B404 mdm4-t_mx_1_random 1 14 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 n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 18 unspecified role unspecified role prey prey physical association physical association Table 1 mdm2-pdi_random_pept 1 15 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 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) 9 unspecified role unspecified role prey prey physical association physical association Table 1 Phage DNA was prepared and the variable region was sequenced. The results showed that 7 of 10 MDMX-selected and 4 of 10 MDM2- selected phages contain the same insert (LTFEHYWAQLTS; Table 1). This peptide was named pDI for peptide dual inhibitor. mdm4-t_mx_4_random 1 10 unspecified role unspecified role prey prey 14 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 n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) physical association physical association Table 1 mdm4-t_mx_9_random 1 14 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 n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 6 unspecified role unspecified role prey prey physical association physical association Table 1 mdm4-pdi_random_pept 1 14 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 n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 9 unspecified role unspecified role prey prey physical association physical association Table 1 Phage DNA was prepared and the variable region was sequenced. The results showed that 7 of 10 MDMX-selected and 4 of 10 MDM2-selected phages contain the same insert (LTFEHYWAQLTS; Table 1). This peptide was named pDI for peptide dual inhibitor. mdm2-t_m2_1_random 1 15 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 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) 11 unspecified role unspecified role prey prey physical association physical association Table 1 mdm2-t_m2_2_random 1 15 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 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) 5 unspecified role unspecified role prey prey physical association physical association Table 1 mdm2-t_m2_3_random-2 1 15 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 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) 16 unspecified role unspecified role prey prey physical association physical association Table 1 mdm2-t_m2_5_random-3 1 12 unspecified role unspecified role prey prey 15 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 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) physical association physical association Table 1 mdm2-t_m2_6_random-4 1 15 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 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) 8 unspecified role unspecified role prey prey physical association physical association Table 1 mdm2-t_m2_9_random-5 1 15 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 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) 17 unspecified role unspecified role prey prey physical association physical association Table 1 tp53-mdm2 2 7 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 ecoli Escherichia coli (strain K12) 15 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 ecoli Escherichia coli (strain K12) physical association physical association 1 A In an ELISA assay, pDI inhibited MDM2-p53 and MDMX-p53 interactions with IC50 of 10 and 100 nmol/L respectively, 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 2 14 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 ecoli Escherichia coli (strain K12) 7 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 ecoli Escherichia coli (strain K12) physical association physical association 1 A In an ELISA assay, pDI inhibited MDM2-p53 and MDMX-p53 interactions with IC50 of 10 and 100 nmol/L respectively, 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 4 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 n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 15 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 in vitro In vitro physical association physical association pDI also inhibited GST-p53 capture of in vitro translated MDM2 and MDMX with different efficiency 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 4 14 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 in vitro In vitro 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 n-terminal sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) physical association physical association pDI also inhibited GST-p53 capture of in vitro translated MDM2 and MDMX with different efficiency 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. thioredoxin_pdi-mdm2 3 15 unspecified role unspecified role prey prey 13 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 F A certain certain sequence position certain certain certain sequence position certain W A certain certain sequence position certain certain certain sequence position certain L A 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 physical association physical association 2 C P0AA25 infection thioredoxin_pdi-mdm4 3 13 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 F A certain certain sequence position certain certain certain sequence position certain W A certain certain sequence position certain certain certain sequence position certain L A 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 14 unspecified role unspecified role prey prey physical association physical association 2 C P0AA25 infection tp53-mdm2-2 3 14 unspecified role unspecified role prey prey 15 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait association association 2 D FLAG-DI expression disrupted MDM2-p53 and MDMX-p53 coprecipitations 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.