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/ Mitochondrial Akt Regulation of Hypoxic Tumor Reprogramming. Mitochondrial Akt Regulation of Hypoxic Tumor Reprogramming. Cancer cell 2016 imex curation Chae YC., Vaira V., Caino MC., Tang HY., Seo JH., Kossenkov AV., Ottobrini L., Martelli C., Lucignani G., Bertolini I., Locatelli M., Bryant KG., Ghosh JC., Lisanti S., Ku B., Bosari S., Languino LR., Speicher DW., Altieri DC. Cancer - Interactions investigated in the context of cancer Further details of PDK proteins were obtained from the authors; PDK1-4 were E.coli expressed, have deleted N-terminal mitochondria targeting sequencse, and have an N-terminal His-tag. daltieri@wistar.org 21-Mar-2017: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro protein kinase assay protein kinase assay western blot western blot Immuno blot Immuno blot Chae YC., Vaira V., Caino MC., Tang HY., Seo JH., Kossenkov AV., Ottobrini L., Martelli C., Lucignani G., Bertolini I., Locatelli M., Bryant KG., Ghosh JC., Lisanti S., Ku B., Bosari S., Languino LR., Speicher DW., Altieri DC. Cancer - Interactions investigated in the context of cancer The phospho-Akt consensus substrate antibody (Akt cons Ab) recognizes peptides and proteins containing phospho-serine/threonine preceded by arginine at the -3 position. There is some preference for peptides that contain phospho-serine/threonine preceded by arginine at both positions -5 and -3. Further details of PDK proteins were obtained from the authors; PDK1-4 were E.coli expressed, have deleted N-terminal mitochondria targeting sequencse, and have an N-terminal His-tag. daltieri@wistar.org Accepted 2017-JAN-20 AT 15:52 GMT AT 15:52 GMT by ORCHARD Cancer cell 2016 imex curation I have added back an Enzyme to EBI-12562450.The GSK3 peptides sold by Cell signalling that I have been able to find are 1)a "blocking peptide" - no species and no sequence given. 2) a GSK-3 Fusion Protein, sequence GPGRRGRRRTSSFAE - chimeric/artificial protein, no species. (Also a full length GSK3-beta human.) there may well be others as the search facility only gives the blocking peptide - if you really want me to enter the chimeric protein, please return to me - not added as of dubious biological validity, and not sure as to which protein used. Fig 3B - I have added 2 Causality statements, but I cannot see that WT "PDK1 then inhibits PDHE1" - only the Kinase Dead mutant, which we cannot enter. Mitochondrial Akt Regulation of Hypoxic Tumor Reprogramming. Mitochondrial Akt Regulation of Hypoxic Tumor Reprogramming. Cancer cell 2016 imex curation Chae YC., Vaira V., Caino MC., Tang HY., Seo JH., Kossenkov AV., Ottobrini L., Martelli C., Lucignani G., Bertolini I., Locatelli M., Bryant KG., Ghosh JC., Lisanti S., Ku B., Bosari S., Languino LR., Speicher DW., Altieri DC. Cancer - Interactions investigated in the context of cancer Further details of PDK proteins were obtained from the authors; PDK1-4 were E.coli expressed, have deleted N-terminal mitochondria targeting sequencse, and have an N-terminal His-tag. daltieri@wistar.org 21-Mar-2017: Contacted by IntAct-Help. Only protein-protein interactions in vitro In vitro protein kinase assay protein kinase assay sequence tag sequence tag identification sequence tag Chae YC., Vaira V., Caino MC., Tang HY., Seo JH., Kossenkov AV., Ottobrini L., Martelli C., Lucignani G., Bertolini I., Locatelli M., Bryant KG., Ghosh JC., Lisanti S., Ku B., Bosari S., Languino LR., Speicher DW., Altieri DC. Cancer - Interactions investigated in the context of cancer Further details of PDK proteins were obtained from the authors; PDK1-4 were E.coli expressed, have deleted N-terminal mitochondria targeting sequencse, and have an N-terminal His-tag. daltieri@wistar.org Accepted 2017-JAN-20 AT 15:52 GMT AT 15:52 GMT by ORCHARD Cancer cell 2016 imex curation I have added back an Enzyme to EBI-12562450.The GSK3 peptides sold by Cell signalling that I have been able to find are 1)a "blocking peptide" - no species and no sequence given. 2) a GSK-3 Fusion Protein, sequence GPGRRGRRRTSSFAE - chimeric/artificial protein, no species. (Also a full length GSK3-beta human.) there may well be others as the search facility only gives the blocking peptide - if you really want me to enter the chimeric protein, please return to me - not added as of dubious biological validity, and not sure as to which protein used. Fig 3B - I have added 2 Causality statements, but I cannot see that WT "PDK1 then inhibits PDHE1" - only the Kinase Dead mutant, which we cannot enter. q15119-1 [Pyruvate dehydrogenase (acetyl-transferring)] kinase isozyme 2, mitochondrial Pyruvate dehydrogenase kinase isoform 2 PDK2 PDHK2 protein protein human Homo sapiens Human MRWVWALLKNASLAGAPKYIEHFSKFSPSPLSMKQFLDFGSSNACEKTSFTFLRQELPVRLANIMKEINLLPDRVLSTPSVQLVQSWYVQSLLDIMEFLDKDPEDHRTLSQFTDALVTIRNRHNDVVPTMAQGVLEYKDTYGDDPVSNQNIQYFLDRFYLSRISIRMLINQHTLIFDGSTNPAHPKHIGSIDPNCNVSEVVKDAYDMAKLLCDKYYMASPDLEIQEINAANSKQPIHMVYVPSHLYHMLFELFKNAMRATVESHESSLILPPIKVMVALGEEDLSIKMSDRGGGVPLRKIERLFSYMYSTAPTPQPGTGGTPLAGFGYGLPISRLYAKYFQGDLQLFSMEGFGTDAVIYLKALSTDSVERLPVYNKSAWRHYQTIQEAGDWCVPSTEPKNTSTYRVS 77DBA03EF922EF03 p31749-1 RAC-alpha serine/threonine-protein kinase AKT1 PKB RAC Protein kinase B Protein kinase B alpha Proto-oncogene c-Akt RAC-PK-alpha protein protein human Homo sapiens Human MSDVAIVKEGWLHKRGEYIKTWRPRYFLLKNDGTFIGYKERPQDVDQREAPLNNFSVAQCQLMKTERPRPNTFIIRCLQWTTVIERTFHVETPEEREEWTTAIQTVADGLKKQEEEEMDFRSGSPSDNSGAEEMEVSLAKPKHRVTMNEFEYLKLLGKGTFGKVILVKEKATGRYYAMKILKKEVIVAKDEVAHTLTENRVLQNSRHPFLTALKYSFQTHDRLCFVMEYANGGELFFHLSRERVFSEDRARFYGAEIVSALDYLHSEKNVVYRDLKLENLMLDKDGHIKITDFGLCKEGIKDGATMKTFCGTPEYLAPEVLEDNDYGRAVDWWGLGVVMYEMMCGRLPFYNQDHEKLFELILMEEIRFPRTLGPEAKSLLSGLLKKDPKQRLGGGSEDAKEIMQHRFFAGIVWQHVYEKKLSPPFKPQVTSETDTRYFDEEFTAQMITITPPDQDDSMECVDSERRPHFPQFSYSASGTA 6EAFF4F8AD436714 pdk4_human [Pyruvate dehydrogenase (acetyl-transferring)] kinase isozyme 4, mitochondrial PDK4 PDHK4 Pyruvate dehydrogenase kinase isoform 4 protein protein human Homo sapiens Human MKAARFVLRSAGSLNGAGLVPREVEHFSRYSPSPLSMKQLLDFGSENACERTSFAFLRQELPVRLANILKEIDILPTQLVNTSSVQLVKSWYIQSLMDLVEFHEKSPDDQKALSDFVDTLIKVRNRHHNVVPTMAQGIIEYKDACTVDPVTNQNLQYFLDRFYMNRISTRMLMNQHILIFSDSQTGNPSHIGSIDPNCDVVAVVQDAFECSRMLCDQYYLSSPELKLTQVNGKFPDQPIHIVYVPSHLHHMLFELFKNAMRATVEHQENQPSLTPIEVIVVLGKEDLTIKISDRGGGVPLRIIDRLFSYTYSTAPTPVMDNSRNAPLAGFGYGLPISRLYAKYFQGDLNLYSLSGYGTDAIIYLKALSSESIEKLPVFNKSAFKHYQMSSEADDWCIPSREPKNLAKEVAM C17B78B6057000E9 q15120-1 [Pyruvate dehydrogenase (acetyl-transferring)] kinase isozyme 3, mitochondrial Pyruvate dehydrogenase kinase isoform 3 PDHK3 PDK3 protein protein human Homo sapiens Human MRLFRWLLKQPVPKQIERYSRFSPSPLSIKQFLDFGRDNACEKTSYMFLRKELPVRLANTMREVNLLPDNLLNRPSVGLVQSWYMQSFLELLEYENKSPEDPQVLDNFLQVLIKVRNRHNDVVPTMAQGVIEYKEKFGFDPFISTNIQYFLDRFYTNRISFRMLINQHTLLFGGDTNPVHPKHIGSIDPTCNVADVVKDAYETAKMLCEQYYLVAPELEVEEFNAKAPDKPIQVVYVPSHLFHMLFELFKNSMRATVELYEDRKEGYPAVKTLVTLGKEDLSIKISDLGGGVPLRKIDRLFNYMYSTAPRPSLEPTRAAPLAGFGYGLPISRLYARYFQGDLKLYSMEGVGTDAVIYLKALSSESFERLPVFNKSAWRHYKTTPEADDWSNPSSEPRDASKYKAKQ EF2415D3F9D8D61E q15118-1 [Pyruvate dehydrogenase (acetyl-transferring)] kinase isozyme 1, mitochondrial PDK1 PDHK1 Pyruvate dehydrogenase kinase isoform 1 protein protein human Homo sapiens Human MRLARLLRGAALAGPGPGLRAAGFSRSFSSDSGSSPASERGVPGQVDFYARFSPSPLSMKQFLDFGSVNACEKTSFMFLRQELPVRLANIMKEISLLPDNLLRTPSVQLVQSWYIQSLQELLDFKDKSAEDAKAIYDFTDTVIRIRNRHNDVIPTMAQGVIEYKESFGVDPVTSQNVQYFLDRFYMSRISIRMLLNQHSLLFGGKGKGSPSHRKHIGSINPNCNVLEVIKDGYENARRLCDLYYINSPELELEELNAKSPGQPIQVVYVPSHLYHMVFELFKNAMRATMEHHANRGVYPPIQVHVTLGNEDLTVKMSDRGGGVPLRKIDRLFNYMYSTAPRPRVETSRAVPLAGFGYGLPISRLYAQYFQGDLKLYSLEGYGTDAVIYIKALSTDSIERLPVYNKAAWKHYNTNHEADDWCVPSREPKDMTTFRSA D14CD594E0EA45A2 odpa_human Pyruvate dehydrogenase E1 component subunit alpha, somatic form, mitochondrial PDHE1-A type I PDHA1 PHE1A protein protein human Homo sapiens Human MRKMLAAVSRVLSGASQKPASRVLVASRNFANDATFEIKKCDLHRLEEGPPVTTVLTREDGLKYYRMMQTVRRMELKADQLYKQKIIRGFCHLCDGQEACCVGLEAGINPTDHLITAYRAHGFTFTRGLSVREILAELTGRKGGCAKGKGGSMHMYAKNFYGGNGIVGAQVPLGAGIALACKYNGKDEVCLTLYGDGAANQGQIFEAYNMAALWKLPCIFICENNRYGMGTSVERAAASTDYYKRGDFIPGLRVDGMDILCVREATRFAAAYCRSGKGPILMELQTYRYHGHSMSDPGVSYRTREEIQEVRSKSDPIMLLKDRMVNSNLASVEELKEIDVEVRKEIEDAAQFATADPEPPLEELGYHIYSSDPPFEVRGANQWIKFKSVS DIP protein P08559 original sequence version: 133 4D685BBE44A92D4B p31751-1 RAC-beta serine/threonine-protein kinase AKT2 Protein kinase Akt-2 Protein kinase B beta RAC-PK-beta protein protein human Homo sapiens Human MNEVSVIKEGWLHKRGEYIKTWRPRYFLLKSDGSFIGYKERPEAPDQTLPPLNNFSVAECQLMKTERPRPNTFVIRCLQWTTVIERTFHVDSPDEREEWMRAIQMVANSLKQRAPGEDPMDYKCGSPSDSSTTEEMEVAVSKARAKVTMNDFDYLKLLGKGTFGKVILVREKATGRYYAMKILRKEVIIAKDEVAHTVTESRVLQNTRHPFLTALKYAFQTHDRLCFVMEYANGGELFFHLSRERVFTEERARFYGAEIVSALEYLHSRDVVYRDIKLENLMLDKDGHIKITDFGLCKEGISDGATMKTFCGTPEYLAPEVLEDNDYGRAVDWWGLGVVMYEMMCGRLPFYNQDHERLFELILMEEIRFPRTLSPEAKSLLAGLLKKDPKQRLGGGPSDAKEVMEHRFFLSINWQDVVQKKLLPPFKPQVTSEVDTRYFDDEFTAQSITITPPDRYDSLGLLELDQRTHFPQFSYSASIRE B18C87A7246BFB24 akt1-pdk1-2 1 4 enzyme enzyme neutral component neutral component n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection 7 enzyme target enzyme target substrate substrate neutral component neutral component n_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation 2C akt1-pdk1-3 1 9 enzyme enzyme neutral component neutral component n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection 7 enzyme target enzyme target substrate substrate neutral component neutral component Q15118-1:p.Thr346Ala mutation disrupting rate mutation disrupting interaction rate mutation disrupting rate certain certain sequence position certain certain certain sequence position certain T A Wed May 02 15:03:01 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'thr346ala' Tue Jun 30 15:45:02 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Thr346Ala' n_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation 2 C/D, 3 (B)/C In the absence of PDK1, PDHE1A was not phosphorylated. akt1-pdk1-4 2 4 enzyme enzyme neutral component neutral component n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection 7 enzyme target enzyme target substrate substrate neutral component neutral component thr346 opthr O-phospho-L-threonine (2S,3R)-2-amino-3-(phosphonooxy)butanoic acid 2-amino-3-hydroxybutanoic acid 3-phosphate 2-azanyl-3-(phosphonooxy)butanoic acid MOD_RES Phosphothreonine O-phospho-L-threonine O-phosphorylated L-threonine O3-phosphothreonine threonine phosphate ester OPThr Phospho Phospho Threonyl Phosphorylation phosphorylationt certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm n_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation 2F, Supp 2C akt2-pdk1-3 2 9 enzyme enzyme neutral component neutral component n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection 7 enzyme target enzyme target substrate substrate neutral component neutral component thr346 opthr O-phospho-L-threonine (2S,3R)-2-amino-3-(phosphonooxy)butanoic acid 2-amino-3-hydroxybutanoic acid 3-phosphate 2-azanyl-3-(phosphonooxy)butanoic acid MOD_RES Phosphothreonine O-phospho-L-threonine O-phosphorylated L-threonine O3-phosphothreonine threonine phosphate ester OPThr Phospho Phospho Threonyl Phosphorylation phosphorylationt certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain n_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation 2F, Supp 2C akt2-pdk1-2 1 9 stimulator stimulator neutral component neutral component n_terminus gst tag glutathione s tranferase tag glutathione S-tranferase tag glutathione S-tranferase tag gst tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection 7 enzyme enzyme neutral component neutral component Q15118-1:p.Thr346Ala mutation decreasing rate mutation decreasing interaction rate mutation decreasing rate certain certain sequence position certain certain certain sequence position certain T A Wed May 02 15:07:12 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'thr346ala' Tue Jun 30 18:32:08 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Thr346Ala' n_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain spofr-sf_9 Spodoptera frugiperda insect cells Fall armyworm sf_9 insect cells Derived from pupal ovarian tissue of the fall armyworm Cloned from the parent cell line, IPLB-SF-21-AE (SF-21) derived from immature ovaries of fall armyworm Spodoptera frugiperda pupae cells are susceptible to Baculovirus infection 8 enzyme target enzyme target substrate substrate neutral component neutral component possible_tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ser293 opser O-phospho-L-serine (2S)-2-amino-3-(phosphonooxy)propanoic acid 2-amino-3-hydroxypropanoic acid 3-phosphate 2-azanyl-3-(phosphonooxy)propanoic acid ACT_SITE Phosphoserine intermediate MOD_RES Phosphoserine O-phospho-L-serine O-phosphonoserine O-phosphorylated L-serine O3-phosphoserine serine phosphate ester OPSer Phospho Phospho Seryl Phosphorylation phosphorylations certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation 3 B/C, Supp 3B. not sure if I have missed something here, but to my eyes the T346A mutant of PDK1 still gives a band showing phosphorylation - despite the text stating "...the T346A PDK1 mutant was ineffective (Figure 3C)". In the absence of PDK1, PDHE1A was not phosphorylated. An increase in PDHE1 phosphorylation was detected only in the presence of Akt2 and PDK1, but not PDK2, PDK3, or PDK4. P31751-1 increases activity of Q15118-1 Q15118-1 is required for phosphorylation of P08559 by Q15118-1 Q15118-1 decreases activity of P08559 akt2-pdha1-1 1 3 enzyme enzyme neutral component neutral component n_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 8 enzyme target enzyme target substrate substrate neutral component neutral component possible_tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ser293 opser O-phospho-L-serine (2S)-2-amino-3-(phosphonooxy)propanoic acid 2-amino-3-hydroxypropanoic acid 3-phosphate 2-azanyl-3-(phosphonooxy)propanoic acid ACT_SITE Phosphoserine intermediate MOD_RES Phosphoserine O-phospho-L-serine O-phosphonoserine O-phosphorylated L-serine O3-phosphoserine serine phosphate ester OPSer Phospho Phospho Seryl Phosphorylation phosphorylations certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation Supp 3B PDK2, PDK3, and PDK4 also phosphorylated PDHE1, but an increase in PDHE1 phosphorylation was only detected when Akt2 was added with PDK1. pdha1-pdk2-1 1 8 enzyme target enzyme target substrate substrate neutral component neutral component ser293 opser O-phospho-L-serine (2S)-2-amino-3-(phosphonooxy)propanoic acid 2-amino-3-hydroxypropanoic acid 3-phosphate 2-azanyl-3-(phosphonooxy)propanoic acid ACT_SITE Phosphoserine intermediate MOD_RES Phosphoserine O-phospho-L-serine O-phosphonoserine O-phosphorylated L-serine O3-phosphoserine serine phosphate ester OPSer Phospho Phospho Seryl Phosphorylation phosphorylations certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm possible_tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) 6 enzyme enzyme neutral component neutral component n_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation Supp 3Bi PDK2, PDK3, and PDK4 also phosphorylated PDHE1, but an increase in PDHE1 phosphorylation was only detected when Akt2 was added with PDK1. pdha1-pdk2-2 1 5 enzyme enzyme neutral component neutral component n_terminus his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain ecoli Escherichia coli (strain K12) 8 enzyme target enzyme target substrate substrate neutral component neutral component possible_tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag n-term range n-terminal range n-term range n-term range n-terminal range n-term range ser293 opser O-phospho-L-serine (2S)-2-amino-3-(phosphonooxy)propanoic acid 2-amino-3-hydroxypropanoic acid 3-phosphate 2-azanyl-3-(phosphonooxy)propanoic acid ACT_SITE Phosphoserine intermediate MOD_RES Phosphoserine O-phospho-L-serine O-phosphonoserine O-phosphorylated L-serine O3-phosphoserine serine phosphate ester OPSer Phospho Phospho Seryl Phosphorylation phosphorylations certain certain sequence position certain certain certain sequence position certain resulting-ptm resulting-ptm resulting ptm resulting-ptm ecoli Escherichia coli (strain K12) phosphorylation phosphorylation reaction phosphorylation Supp 3Bii PDK2, PDK3, and PDK4 also phosphorylated PDHE1, but an increase in PDHE1 phosphorylation was only detected when Akt2 was added with PDK1.