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 Adi3 is a Pdk1-interacting AGC kinase that negatively regulates plant cell death. Adi3 is a Pdk1-interacting AGC kinase that negatively regulates plant cell death. EMBO J. (0261-4189) 2006 imex curation Devarenne TP., Ekengren SK., Pedley KF., Martin GB. 2009-01-08 Only protein-protein interactions in vitro In vitro anti tag coip anti tag coimmunoprecipitation anti tag coip nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Devarenne TP., Ekengren SK., Pedley KF., Martin GB. EMBO J. (0261-4189) 2006 2009-01-08 Only protein-protein interactions imex curation Adi3 is a Pdk1-interacting AGC kinase that negatively regulates plant cell death. Adi3 is a Pdk1-interacting AGC kinase that negatively regulates plant cell death. EMBO J. (0261-4189) 2006 imex curation Devarenne TP., Ekengren SK., Pedley KF., Martin GB. 2009-01-08 Only protein-protein interactions yeast Saccharomyces cerevisiae Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence Devarenne TP., Ekengren SK., Pedley KF., Martin GB. EMBO J. (0261-4189) 2006 2009-01-08 Only protein-protein interactions imex curation Adi3 is a Pdk1-interacting AGC kinase that negatively regulates plant cell death. Adi3 is a Pdk1-interacting AGC kinase that negatively regulates plant cell death. EMBO J. (0261-4189) 2006 imex curation Devarenne TP., Ekengren SK., Pedley KF., Martin GB. 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 Devarenne TP., Ekengren SK., Pedley KF., Martin GB. EMBO J. (0261-4189) 2006 2009-01-08 Only protein-protein interactions imex curation q5i6e8_sollc Non-specific serine/threonine protein kinase Pdk1 protein protein sollc Solanum lycopersicum (Tomato) Lycopersicon esculentum Tomato MLALVGGEGDMEQEFDAKLKIQNNSANTQRSKSFAFRAPQENFTIQDFELGKIYGVGSYSKVVRAKKKDTANVYALKIMDKKFITKENKTAYVKLERIVLDQLDHPGVVRLFFTFQDTFSLYMALESCEGGELFDQITRKGRLSEDEARFYAAEVVDALEYIHSMGLIHRDIKPENLLLTSDGHIKIADFGSVKPMQDSRITVLPNAASDDKACTFVGTAAYVPPEVLNSSPATFGNDLWALGCTLYQMLSGTSPFKDASEWLIFQRIIARDIRFPNYFSNEARDIIDQLLDVDPSRRPGAGPDGYASLKNHPFFSGIDWENLRLQTPPRLAMEPKAPSTHSSGDEQDPSWNPSHIGDGSVRPNDGNGAAASVSEAGNSITRLASIDSFDSKWKQFLDPGESVLMISMVKKLQKLTSKKVQLILTNKPKLIYVDPSKLVIKGNIIWSDNPNDLSIQVTSPSQFKICTPKKVMSFEDAKNRAQQWKKAIEALQNR mint D90C34DDD45FC87F p93215_sollc pto 101268866 protein protein sollc Solanum lycopersicum (Tomato) Lycopersicon esculentum Tomato MGSKYSKATNSISDASNSFESYRFPLEDLEEATNNFDDKFFIGEGAFGKVYKGVLRDGTKVALKRQNRDSRQGIEEFGTEIGILSRRSHPHLVSLIGYCDERNEMVLIYDYMENGNLKSHLTGSDLPSMSWEQRLEICIGAARGLHYLHTNGVMHRDVKSSNILLDENFVPKITDFGLSKTRPQLYQTTDVKGTFGYIDPEYFIKGRLTEKSDVYSFGVVLFEVLCARSAMVQSLPREMVNLAEWAVESHNNGQLEQIVDPNLADKIRPESLRKFGETAVKCLALSSEDRPSMGDVLWKLEYALRLQESVI mint B85B89A8380499E7 q5i6e9_sollc Adi3 protein protein sollc Solanum lycopersicum (Tomato) Lycopersicon esculentum Tomato MERIPEVRESTRQFPIGAKVAHTFSTSKKEVGIRGFRDFDLAIPIQTWKGKTSYQEEEDLMVDAGTIKRSDDSLEDSGSTSFHGASHPPEPVDTDLMRPVYVPIGQNKADGKCLVKNVSLKGPFLDDLSIRMPNVKPSPSLLSPAESLVEEPNDLGVISSPFTVPRPSQNTETSLPPDSEEKECIWDASLPPSGNVSPLSSIDSTGVVRSMSIVNSCTSTYRSDVLMSDGMLSVDRNYESTKGSIRGDSLESGKTSLSRASDSSGLSDDSNWSNITGSANKPHKGNDPRWKAILAIRARDGILGMSHFKLLKRLGCGDIGSVYLSELSGTRCYFAMKVMDKASLASRKKLTRAQTEREILQLLDHPFLPTLYTHFETDRFSCLVMEYCPGGDLHTLRQRQPGKHFSEYAARFYAAEVLLALEYLHMLGVVYRDLKPENVLVRDDGHIMLSDFDLSLRCAVSPTLIRISSDDPSKRGAAFCVQPACIEPTTVCMQPACFLPRLFPQKSKKKTPKPRADSGFQANSMPELVAEPTSARSMSFVGTHEYLAPEIIKGEGHGSAVDWWTFGIFLHELLYGKTPFKGSGNRATLFNVVGQQLKFPDSPATSYASRDLIRGLLVKEPQNRLGVKRGATEIKQHPFFEGVNWALIRCSTPPEVPRPVEPDYPAKYGQVNPVGVGNTSKRVVGADAKSGGKYLDFEFF 87C79B60D952B6B1 pdk1-adi3 2 4 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 tagged molecule tag 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 unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell tagged molecule tag tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 physical association physical association F2B EMBO-Journal In the Y2H assay, Pdk1 did not interact with Pto or AvrPto, but did interact with Adi3 irrespective of the kinase activity of Adi3 or Pdk1 (Figure 2B). mint adi3-pdk1 3 4 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined enzyme enzyme neutral component neutral component purified purified mbp tagged mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp 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 phosphorylated phosres phosphorylated residue PhosRes Phospho Phosphorylation Phosphorylation (O of Serine, Threonine, Tyrosine and Aspartate, N epsilon of Lysine) certain certain sequence position certain certain certain sequence position certain range MINT-2740214: original sequence: 539-539 : VAEPTSARSMSFVGTHEYLA undefined mutation mutation decreasing interaction mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Wed Jul 06 16:23:45 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 decreasing' Phe698Ala range MINT-2740220: original sequence: 698-698 : AKSGGKYLDFEF mbp tagged mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Q5I6E9:p.Phe697Ala mutation decreasing interaction mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain F A Tue Jul 12 10:21:34 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation decreasing' Wed May 02 18:24:41 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'phe697ala' Tue Jun 30 17:40:46 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Phe697Ala' range MINT-2740217: original sequence: 697-697 : DAKSGGKYLDFEF undefined mutation mutation decreasing interaction mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain Wed Jul 06 16:24:16 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 decreasing' Glu700Ala range MINT-2740223: original sequence: 700-700 : SGGKYLDFEF Stoichiometry: 1.0 phosphorylation phosphorylation reaction phosphorylation F3B F2C F2E F2F EMBO-Journal Mutation of S539 to Asp, simulating a phosphorylation event, greatly increases the autophosphorylation activity of Adi3 (Figure 1A), suggesting that phosphorylation of S539 by Pdk1 is required for activation of Adi3. MBP-Pdk1 phosphorylated MBP-Adi3 but it phosphorylated MBP-Adi3S539A at a much reduced level (Figure 2C). MBP-Pdk1 phosphorylation of MBP-Adi3K337Q followed by phosphoamino acid analysis indicates that only Ser residues are phosphorylated on Adi3 by Pdk1 (Figure 2E). Combining all three substitutions gave the greatest loss of Pdk1 phosphorylation of Adi3 (Figure 3B). Taken together, these data indicate that Pdk1 phosphorylates Adi3 at S539 (Figure 2C, E, and F) mint adi3-adi3 3 6 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined enzyme enzyme self self purified purified Q5I6E9:p.Glu698Ala mutation decreasing interaction mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain E A Tue Jul 12 10:23:00 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation decreasing' Wed May 02 16:07:54 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'glu698ala' Tue Jun 30 16:56:08 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Glu698Ala' range MINT-2740323: original sequence: 698-698 : AKSGGKYLDFEF Q5I6E9:p.Phe697Ala mutation decreasing interaction mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain F A Tue Jul 12 10:58:21 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation decreasing' Wed May 02 19:13:26 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'phe697ala' Tue Jun 30 17:52:40 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Phe697Ala' range MINT-2740320: original sequence: 697-697 : DAKSGGKYLDFEF Q5I6E9:p.Phe700Ala mutation decreasing interaction mutation disrupting mutation disrupting interaction mutation disrupting mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain F A Tue Jul 12 10:22:56 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation decreasing' Wed May 02 19:41:14 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'phe700ala' Tue Jun 30 19:53:43 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Phe700Ala' range MINT-2740326: original sequence: 700-700 : SGGKYLDFEF 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 self self purified purified mbp tagged mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 phosphorylation phosphorylation reaction phosphorylation F1A F3D EMBO-Journal e examined the possible autophosphorylation activity of MBP-Adi3. MBP-Adi3 showed autophosphorylation activity, whereas MBP-Adi3K337Q, which has a substitution at the ATP-binding site, did not (Figure 1A). Mutation of F697, E698, and F700 to Ala, singly or in combination, completely eliminated Adi3 autophosphorylation (Figure 3D). mint adi3-pto 3 5 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined enzyme enzyme neutral component neutral component purified purified mbp tagged mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp 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 mbp tagged mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 phosphorylation phosphorylation reaction phosphorylation F1C EMBO-Journal When MBP-Adi3 was incubated with MBP-PtoD164N, phosphorylation of PtoD164N was not seen regardless of the presence of AvrPto (Figure 1C, lanes 7 and 8). These data demonstrate that Adi3 is a substrate for Pto. mint pdk1-pdk1 3 4 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined enzyme enzyme self self purified purified Q5I6E8:p.Lys77Gln 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 Q Tue Jul 12 10:23:44 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation disrupting' Wed May 02 17:45:54 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'lys77gln' Tue Jun 30 20:04:09 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Lys77Gln' range MINT-2740434: original sequence: 77-77 : KKDTANVYALKIMDKKFITK mbp tagged mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 4 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined enzyme target enzyme target substrate substrate self self purified purified mbp tagged mbp tag maltose binding protein tag maltose binding protein tag maltose binding protein tag mbp tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 1.0 phosphorylation phosphorylation reaction phosphorylation F2A EMBO-Journal We found that it is an active protein kinase in autophosphorylation assays by using kinase-active MBP-Pdk1 and kinase-inactive MBP-Pdk1K77Q (subdomain II conserved Lys) proteins (Figure 2A). mint pdk1-adi3-1 1 4 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role bait bait purified purified 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 6 nucleotide sequence nucleotide sequence identification sequence cloning sequence cloning nucleotide sequence predetermined predetermined participant predetermined unspecified role unspecified role prey prey purified purified Stoichiometry: 1.0 direct interaction direct interaction F3C F2D EMBO-Journal MBP-Adi3 mutant proteins co-immunoprecipitated in vitro with Pdk1-FLAG in equal amounts (Figure 2D). mint