MINT MINT, Dpt of Biology, University of Rome Tor Vergata MINT http://mint.bio.uniroma2.it/mint url>http://mint.bio.uniroma2.it/mint/ https://mint.bio.uniroma2.it MINT-[0-9]+ https://mint.bio.uniroma2.it/index.php/results-interactions/?id=${ac} The Molecular INTeraction database (MINT) is a relational database designed to store interactions between biological molecules. http://mint.bio.uniroma2.it 14-3-3gamma binds to MDMX that is phosphorylated by UV-activated Chk1, resulting in p53 activation. 14-3-3gamma binds to MDMX that is phosphorylated by UV-activated Chk1, resulting in p53 activation. EMBO J. (0261-4189) 2006 imex curation Jin Y., Dai MS., Lu SZ., Xu Y., Luo Z., Zhao Y., Lu H. Virus - Publications including interactions involving viral proteins 2009-01-08 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 predetermined predetermined participant predetermined Jin Y., Dai MS., Lu SZ., Xu Y., Luo Z., Zhao Y., Lu H. Virus - Publications including interactions involving viral proteins EMBO J. (0261-4189) 2006 2009-01-08 Only protein-protein interactions imex curation 14-3-3gamma binds to MDMX that is phosphorylated by UV-activated Chk1, resulting in p53 activation. 14-3-3gamma binds to MDMX that is phosphorylated by UV-activated Chk1, resulting in p53 activation. EMBO J. (0261-4189) 2006 imex curation Jin Y., Dai MS., Lu SZ., Xu Y., Luo Z., Zhao Y., Lu H. Virus - Publications including interactions involving viral proteins 2009-01-08 Only protein-protein interactions in vitro In vitro pull down pull down nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Jin Y., Dai MS., Lu SZ., Xu Y., Luo Z., Zhao Y., Lu H. Virus - Publications including interactions involving viral proteins EMBO J. (0261-4189) 2006 2009-01-08 Only protein-protein interactions imex curation 14-3-3gamma binds to MDMX that is phosphorylated by UV-activated Chk1, resulting in p53 activation. 14-3-3gamma binds to MDMX that is phosphorylated by UV-activated Chk1, resulting in p53 activation. EMBO J. (0261-4189) 2006 imex curation Jin Y., Dai MS., Lu SZ., Xu Y., Luo Z., Zhao Y., Lu H. Virus - Publications including interactions involving viral proteins 2009-01-08 Only protein-protein interactions human-293 Homo sapiens transformed primary embryonal kidney cells Human 293 Transformed human primary embryonal kidney cells. The Cell type named 293 is synonymous with HEK-293. anti tag coip anti tag coimmunoprecipitation anti tag coip nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Jin Y., Dai MS., Lu SZ., Xu Y., Luo Z., Zhao Y., Lu H. Virus - Publications including interactions involving viral proteins EMBO J. (0261-4189) 2006 2009-01-08 Only protein-protein interactions imex curation 14-3-3gamma binds to MDMX that is phosphorylated by UV-activated Chk1, resulting in p53 activation. 14-3-3gamma binds to MDMX that is phosphorylated by UV-activated Chk1, resulting in p53 activation. EMBO J. (0261-4189) 2006 imex curation Jin Y., Dai MS., Lu SZ., Xu Y., Luo Z., Zhao Y., Lu H. Virus - Publications including interactions involving viral proteins 2009-01-08 Only protein-protein interactions in vitro In vitro protein kinase assay protein kinase assay nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Jin Y., Dai MS., Lu SZ., Xu Y., Luo Z., Zhao Y., Lu H. Virus - Publications including interactions involving viral proteins EMBO J. (0261-4189) 2006 2009-01-08 Only protein-protein interactions imex curation 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 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 1433g_human 14-3-3 protein gamma YWHAG Protein kinase C inhibitor protein 1 protein protein human Homo sapiens Human MVDREQLVQKARLAEQAERYDDMAAAMKNVTELNEPLSNEERNLLSVAYKNVVGARRSSWRVISSIEQKTSADGNEKKIEMVRAYREKIEKELEAVCQDVLSLLDNYLIKNCSETQYESKVFYLKMKGDYYRYLAEVATGEKRATVVESSEKAYSEAHEISKEHMQPTHPIRLGLALNYSVFYYEIQNAPEQACHLAKTAFDDAIAELDTLNEDSYKDSTLIMQLLRDNLTLWTSDQQDDDGGEGNN DIP protein P61981 original sequence version: 126 DIP protein P61981 original sequence version: 135 B0D16C6DE1F4455D 1433s_human 14-3-3 protein sigma Stratifin Epithelial cell marker protein 1 HME1 SFN protein protein human Homo sapiens Human MERASLIQKAKLAEQAERYEDMAAFMKGAVEKGEELSCEERNLLSVAYKNVVGGQRAAWRVLSSIEQKSNEEGSEEKGPEVREYREKVETELQGVCDTVLGLLDSHLIKEAGDAESRVFYLKMKGDYYRYLAEVATGDDKKRIIDSARSAYQEAMDISKKEMPPTNPIRLGLALNFSVFHYEIANSPEEAISLAKTTFDEAMADLHTLSEDSYKDSTLIMQLLRDNLTLWTADNAGEEGGEAPQEPQS DIP protein P31947 original sequence version: 84 7F4B44E3AA59ECE6 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 q3yba8_human protein protein human Homo sapiens Human MEKTELIQKAKLAERYDDMATCMKAGTKQGAELSKEERNLLSVAYKNVVGAAGPPGASSGASSRIPTPPTRSCS mint homomint 57F6F5655A97CEA8 ywhag-mdm4-1 1 7 polyclonal western polyclonal antibody western blot polyclonal western predetermined predetermined participant predetermined unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Stoichiometry: 1.0 9 monoclonal western monoclonal antibody western blot monoclonal western predetermined predetermined participant predetermined unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Stoichiometry: 1.0 physical association physical association F2C EMBO-Journal Endogenous MDMX and 14-3-3gamma were coimmunoprecipitated with the anti-14-3-3gamma or anti-MDMX, but not anti-His, antibody from 293 cells (Figure 2C). homomint domino mint mdm4-ywhag 1 9 monoclonal western monoclonal antibody western blot monoclonal western predetermined predetermined participant predetermined unspecified role unspecified role bait bait living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Stoichiometry: 1.0 7 polyclonal western polyclonal antibody western blot polyclonal western predetermined predetermined participant predetermined unspecified role unspecified role prey prey living cell living cell physiological level physiological level endogenous level endogenous endogenous endogenous level Stoichiometry: 1.0 physical association physical association F2C EMBO-Journal Endogenous MDMX and 14-3-3gamma were coimmunoprecipitated with the anti-14-3-3gamma or anti-MDMX, but not anti-His, antibody from 293 cells (Figure 2C). homomint domino mint ywhag-mdm4-2 3 7 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell P61981:p.Lys50Glu mutation disrupting interaction mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain K E Tue Jul 12 10:15:08 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation disrupting' Wed May 02 18:54:44 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys50glu' Tue Jun 30 16:43:09 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys50Glu' range MINT-1957809: original sequence: 50-50 : EERNLLSVAYKNVVGARRSS flag tagged 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 Stoichiometry: 1.0 9 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell O15151:p.Ser367Gly mutation disrupting interaction mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain S G Tue Jul 12 10:53:59 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation disrupting' Wed May 02 14:49:21 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser367gly' Tue Jun 30 17:28:54 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser367Gly' range MINT-1957825: original sequence: 367-367 : NDVPDCRRTISAPVVRPKDA undefined mutation mutation disrupting interaction mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Wed Jul 06 16:52:40 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'mutation disrupting' Ser369Gly range MINT-1957831: original sequence: 369-369 : VPDCRRTISAPVVRPKDAYI myc tagged myc tagged myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F2A F3C F3D EMBO-Journal MDMX bound to 14-3-3gamma more efficiently than to the sigma, tau, alt epsilon, beta, or zeta isoforms, as tested using IP with anti-Flag (Figure 2A). Mutation of either S367 or P369 in MDMX completely abolished the interaction between MDMX and 14-3-3gamma (Figure 3C). In an experiment similar to that shown in Figure 3C, except the K50E mutant, we found that this mutant failed to bind MDMX in cells (Figure 3D). homomint domino mint q3yba8-mdm4-2 3 10 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell flag tagged flag tagged 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 Stoichiometry: 1.0 9 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell myc tagged myc tagged myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F2A EMBO-Journal MDMX bound to 14-3-3gamma more efficiently than to the sigma, tau, alt epsilon, beta, or zeta isoforms, as tested using IP with anti-Flag (Figure 2A). homomint domino mint sfn-mdm4 3 9 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell myc tagged myc tagged myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 8 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell flag tagged flag tagged 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 Stoichiometry: 1.0 physical association physical association F2A EMBO-Journal MDMX bound to 14-3-3gamma more efficiently than to the sigma, tau, alt epsilon, beta, or zeta isoforms, as tested using IP with anti-Flag (Figure 2A). homomint domino mint mdm4-ywhag-1 3 7 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell flag tagged flag tagged 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 Stoichiometry: 1.0 9 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell gfp tagged gfp tagged gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F2B EMBO-Journal MDMX bound to 14-3-3gamma more efficiently than to the sigma, tau, alt epsilon, beta, or zeta isoforms, as tested using IP with anti-Flag (Figure 2A) and anti-GFP (Figure 2B). homomint domino mint mdm4-ywhag-2 3 9 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell gfp tagged gfp tagged gfp tag green fluorescent protein tag GFP green fluorescent protein GFP green fluorescent protein gfp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 7 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell flag tagged flag tagged 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 Stoichiometry: 1.0 physical association physical association SF1 EMBO-Journal MDMX bound to 14-3-3gamma more efficiently than to the sigma, tau, alt epsilon, beta, or zeta isoforms, as tested using IP with anti-Flag (Figure 2A) and anti-GFP (Figure 2B) (see Supplementary Figure S1 for 14-3-3alt epsilon; data for beta and zeta isoforms are not shown). homomint domino mint ywhag-mdm4-3 2 7 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified gst tagged 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 Stoichiometry: 1.0 9 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain range MINT-1958286: original sequence: 359-373 : VPDCRRTISAPVVR his tagged his tagged 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 Stoichiometry: 1.0 direct interaction direct interaction F3A F3B EMBO-Journal (Figure 3A): Indeed, MDMX bound to GST-14-3-3gamma (lane 2), but not GST alone (lane 1). This binding was reduced by a 15-mer peptide that contains the serine-phosphorylated 14-3-3-binding consensus sequence RSASpEP, but not by its nonphosphorylated counterpart, in a dose-dependent manner (Figure 3A). The same result was obtained when a serine-phosphorylated 15-mer peptide containing the sequence 363RRTISpAP369 derived from MDMX was used under the same experimental setting (Figure 3B). This binding was reduced when a serine-phosphorylated 15-mer peptide containing the sequence 363RRTISpAP369 derived from MDMX was used under the same experimental setting (Figure 3B). homomint domino mint chek1-mdm4 4 5 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined enzyme enzyme neutral component neutral component purified purified gst tagged gst tagged 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 Stoichiometry: 1.0 9 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined enzyme target enzyme target substrate substrate neutral component neutral component purified purified phosphorylated phosphorylated residue phosres phosphorylated residue PhosRes Phospho Phosphorylation Phosphorylation (O of Serine, Threonine, Tyrosine and Aspartate, N epsilon of Lysine) western blot western blot Immuno blot Immuno blot certain certain sequence position certain certain certain sequence position certain range MINT-1958323: original sequence: 367-367 : NDVPDCRRTISAPVVRPKDA his tagged his tagged 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 Stoichiometry: 1.0 phosphorylation phosphorylation reaction phosphorylation F6B EMBO-Journal Using the purified antibody, we found that it detected the phosphorylated species of MDMX by Chk1, but not by its KD mutant, in vitro (Figure 6B). homomint mint chek1-tp53 4 5 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined enzyme enzyme neutral component neutral component purified purified gst tagged gst tagged 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 Stoichiometry: 1.0 6 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined enzyme target enzyme target substrate substrate neutral component neutral component purified purified his tagged his tagged 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 Stoichiometry: 1.0 phosphorylation phosphorylation reaction phosphorylation EMBO-Journal SF4A Our in vitro kinase assay showed that UNC-01 effectively inhibited Chk1 activity on p53. In all, 300 nM of the inhibitor led to 80% reduction of p53 phosphorylation (Supplementary Figure S4a). homomint mint