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 A thermodynamic switch modulates abscisic acid receptor sensitivity. A thermodynamic switch modulates abscisic acid receptor sensitivity. EMBO J. (0261-4189) 2011 imex curation Dupeux F., Santiago J., Betz K., Twycross J., Park SY., Rodriguez L., Gonzalez-Guzman M., Jensen MR., Krasnogor N., Blackledge M., Holdsworth M., Cutler SR., Rodriguez PL., Márquez JA. 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 predetermined predetermined participant predetermined Dupeux F., Santiago J., Betz K., Twycross J., Park SY., Rodriguez L., Gonzalez-Guzman M., Jensen MR., Krasnogor N., Blackledge M., Holdsworth M., Cutler SR., Rodriguez PL., Márquez JA. EMBO J. (0261-4189) 2011 Only protein-protein interactions imex curation A thermodynamic switch modulates abscisic acid receptor sensitivity. A thermodynamic switch modulates abscisic acid receptor sensitivity. EMBO J. (0261-4189) 2011 imex curation Dupeux F., Santiago J., Betz K., Twycross J., Park SY., Rodriguez L., Gonzalez-Guzman M., Jensen MR., Krasnogor N., Blackledge M., Holdsworth M., Cutler SR., Rodriguez PL., Márquez JA. Only protein-protein interactions in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Dupeux F., Santiago J., Betz K., Twycross J., Park SY., Rodriguez L., Gonzalez-Guzman M., Jensen MR., Krasnogor N., Blackledge M., Holdsworth M., Cutler SR., Rodriguez PL., Márquez JA. EMBO J. (0261-4189) 2011 Only protein-protein interactions imex curation A thermodynamic switch modulates abscisic acid receptor sensitivity. A thermodynamic switch modulates abscisic acid receptor sensitivity. EMBO J. (0261-4189) 2011 imex curation Dupeux F., Santiago J., Betz K., Twycross J., Park SY., Rodriguez L., Gonzalez-Guzman M., Jensen MR., Krasnogor N., Blackledge M., Holdsworth M., Cutler SR., Rodriguez PL., Márquez JA. Only protein-protein interactions in vitro In vitro nmr nuclear magnetic resonance NMR NMR nmr predetermined predetermined participant predetermined Dupeux F., Santiago J., Betz K., Twycross J., Park SY., Rodriguez L., Gonzalez-Guzman M., Jensen MR., Krasnogor N., Blackledge M., Holdsworth M., Cutler SR., Rodriguez PL., Márquez JA. EMBO J. (0261-4189) 2011 Only protein-protein interactions imex curation A thermodynamic switch modulates abscisic acid receptor sensitivity. A thermodynamic switch modulates abscisic acid receptor sensitivity. EMBO J. (0261-4189) 2011 imex curation Dupeux F., Santiago J., Betz K., Twycross J., Park SY., Rodriguez L., Gonzalez-Guzman M., Jensen MR., Krasnogor N., Blackledge M., Holdsworth M., Cutler SR., Rodriguez PL., Márquez JA. Only protein-protein interactions in vitro In vitro itc isothermal titration calorimetry ITC ITC itc predetermined predetermined participant predetermined Dupeux F., Santiago J., Betz K., Twycross J., Park SY., Rodriguez L., Gonzalez-Guzman M., Jensen MR., Krasnogor N., Blackledge M., Holdsworth M., Cutler SR., Rodriguez PL., Márquez JA. EMBO J. (0261-4189) 2011 Only protein-protein interactions imex curation A thermodynamic switch modulates abscisic acid receptor sensitivity. A thermodynamic switch modulates abscisic acid receptor sensitivity. EMBO J. (0261-4189) 2011 imex curation Dupeux F., Santiago J., Betz K., Twycross J., Park SY., Rodriguez L., Gonzalez-Guzman M., Jensen MR., Krasnogor N., Blackledge M., Holdsworth M., Cutler SR., Rodriguez PL., Márquez JA. Only protein-protein interactions in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column predetermined predetermined participant predetermined Dupeux F., Santiago J., Betz K., Twycross J., Park SY., Rodriguez L., Gonzalez-Guzman M., Jensen MR., Krasnogor N., Blackledge M., Holdsworth M., Cutler SR., Rodriguez PL., Márquez JA. EMBO J. (0261-4189) 2011 Only protein-protein interactions imex curation p2c16_arath Protein phosphatase 2C 16 HAB1 P2C-HA Protein phosphatase 2C HAB1 AtP2C-HA Protein HYPERSENSITIVE TO ABA 1 F28P22.4 At1g72770 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MEEMTPAVAMTLSLAANTMCESSPVEITQLKNVTDAADLLSDSENQSFCNGGTECTMEDVSELEEVGEQDLLKTLSDTRSGSSNVFDEDDVLSVVEDNSAVISEGLLVVDAGSELSLSNTAMEIDNGRVLATAIIVGESSIEQVPTAEVLIAGVNQDTNTSEVVIRLPDENSNHLVKGRSVYELDCIPLWGTVSIQGNRSEMEDAFAVSPHFLKLPIKMLMGDHEGMSPSLTHLTGHFFGVYDGHGGHKVADYCRDRLHFALAEEIERIKDELCKRNTGEGRQVQWDKVFTSCFLTVDGEIEGKIGRAVVGSSDKVLEAVASETVGSTAVVALVCSSHIVVSNCGDSRAVLFRGKEAMPLSVDHKPDREDEYARIENAGGKVIQWQGARVFGVLAMSRSIGDRYLKPYVIPEPEVTFMPRSREDECLILASDGLWDVMNNQEVCEIARRRILMWHKKNGAPPLAERGKGIDPACQAAADYLSMLALQKGSKDNISIIVIDLKAQRKFKTRT DIP protein Q9CAJ0 original sequence version: 64 DIP protein Q9CAJ0-1 original sequence version: 62 DIP protein Q9CAJ0-1 original sequence version: 77 DIP protein Q9CAJ0 original sequence version: 62 DIP protein Q9CAJ0 original sequence version: 65 DIP protein Q9CAJ0-1 original sequence version: 66 DIP protein Q9CAJ0 original sequence version: 81 DIP protein Q9CAJ0-1 original sequence version: 86 DIP protein Q9CAJ0-1 original sequence version: 108 DIP protein Q9CAJ0 original sequence version: 112 DIP protein Q9CAJ0-1 original sequence version: 81 B001BAF0DD8333E1 pyl5_arath Abscisic acid receptor PYL5 At5g05440 PYL5 ABIP3 RCAR8 PYR1-like protein 5 ABI1-binding protein 3 Regulatory components of ABA receptor 8 K18I23.25 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MRSPVQLQHGSDATNGFHTLQPHDQTDGPIKRVCLTRGMHVPEHVAMHHTHDVGPDQCCSSVVQMIHAPPESVWALVRRFDNPKVYKNFIRQCRIVQGDGLHVGDLREVMVVSGLPAVSSTERLEILDEERHVISFSVVGGDHRLKNYRSVTTLHASDDEGTVVVESYIVDVPPGNTEEETLSFVDTIVRCNLQSLARSTNRQ DIP protein Q9FLB1 original sequence version: 46 DIP protein Q9FLB1 original sequence version: 51 5A446F5E40D5AF11 pyl1_arath Abscisic acid receptor PYL1 At5g46790 PYL1 RCAR12 PYR1-like protein 1 ABI1-binding protein 6 Regulatory components of ABA receptor 9 MZA15.21 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MANSESSSSPVNEEENSQRISTLHHQTMPSDLTQDEFTQLSQSIAEFHTYQLGNGRCSSLLAQRIHAPPETVWSVVRRFDRPQIYKHFIKSCNVSEDFEMRVGCTRDVNVISGLPANTSRERLDLLDDDRRVTGFSITGGEHRLRNYKSVTTVHRFEKEEEEERIWTVVLESYVVDVPEGNSEEDTRLFADTVIRLNLQKLASITEAMNRNNNNNNSSQVR DIP protein Q8VZS8 original sequence version: 33 DIP protein Q8VZS8 original sequence version: 38 DIP protein Q8VZS8 original sequence version: 34 DIP protein Q8VZS8 original sequence version: 39 B64B1A52C43072C2 pyl6_arath Abscisic acid receptor PYL6 T3G21 At2g40330 PYL6 ABIP5 RCAR9 PYR1-like protein 6 ABI1-binding protein 5 Regulatory components of ABA receptor 9 T7M7.15 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MPTSIQFQRSSTAAEAANATVRNYPHHHQKQVQKVSLTRGMADVPEHVELSHTHVVGPSQCFSVVVQDVEAPVSTVWSILSRFEHPQAYKHFVKSCHVVIGDGREVGSVREVRVVSGLPAAFSLERLEIMDDDRHVISFSVVGGDHRLMNYKSVTTVHESEEDSDGKKRTRVVESYVVDVPAGNDKEETCSFADTIVRCNLQSLAKLAENTSKFS DIP protein Q8S8E3 original sequence version: 35 DIP protein Q8S8E3 original sequence version: 40 8B9D3F7BD9E2860D pyl8_arath Abscisic acid receptor PYL8 PYL8 ABIP1 RCAR3 PYR1-like protein 8 ABI1-binding protein 1 Regulatory components of ABA receptor 3 MFH8.10 At5g53160 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MEANGIENLTNPNQEREFIRRHHKHELVDNQCSSTLVKHINAPVHIVWSLVRRFDQPQKYKPFISRCVVKGNMEIGTVREVDVKSGLPATRSTERLELLDDNEHILSIRIVGGDHRLKNYSSIISLHPETIEGRIGTLVIESFVVDVPEGNTKDETCYFVEALIKCNLKSLADISERLAVQDTTESRV D3436ADAC890AEED pyr1_arath Abscisic acid receptor PYR1 PYR1 ABIP6 At4g17870 RCAR11 Protein PYRABACTIN RESISTANCE 1 ABI1-binding protein 6 Regulatory components of ABA receptor 11 T6K21.50 protein protein arath Arabidopsis thaliana (Mouse-ear cress) Mouse-ear cress MPSELTPEERSELKNSIAEFHTYQLDPGSCSSLHAQRIHAPPELVWSIVRRFDKPQTYKHFIKSCSVEQNFEMRVGCTRDVIVISGLPANTSTERLDILDDERRVTGFSIIGGEHRLTNYKSVTTVHRFEKENRIWTVVLESYVVDMPEGNSEDDTRMFADTVVKLNLQKLATVAEAMARNSGDGSGSQVT DIP protein O49686 original sequence version: 50 DIP protein O49686 original sequence version: 63 DIP protein O49686 original sequence version: 54 DIP protein O49686 original sequence version: 92 AD35DA240E286B55 pyr1-4 5 Pyr1 11 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified Stoichiometry: 2.0 direct interaction direct interaction f2 mint hab1-pyr1-1 5 HAB1 6 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain Pyr1 11 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified direct interaction direct interaction mint all receptors tested form stable complexes with DNHAB1 in the presence of ABA at 1:1 stoichiometry, as indicated by the apparent Mr of the complexes (right panels in Figure 2; Supplementary Figure S1). Importantly, PYR1 and PYL1 do not interact with PP2C in the absence of ABA, while PYL5, PYL6 and PYL8 show partial interactions (Figure 2; Supplementary Figure S1). f2 hab1-pyl1 5 Pyl1 8 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified HAB1 6 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain direct interaction direct interaction sf1 mint all receptors tested form stable complexes with DNHAB1 in the presence of ABA at 1:1 stoichiometry, as indicated by the apparent Mr of the complexes (right panels in Figure 2; Supplementary Figure S1). Importantly, PYR1 and PYL1 do not interact with PP2C in the absence of ABA, while PYL5, PYL6 and PYL8 show partial interactions (Figure 2; Supplementary Figure S1). pyl1-1 5 Pyl1 8 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified Stoichiometry: 2.0 direct interaction direct interaction sf1 mint hab1-pyl8-2 5 HAB1 6 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain Pyl8 10 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified direct interaction direct interaction sf1 mint all receptors tested form stable complexes with DNHAB1 in the presence of ABA at 1:1 stoichiometry, as indicated by the apparent Mr of the complexes (right panels in Figure 2; Supplementary Figure S1). Importantly, PYR1 and PYL1 do not interact with PP2C in the absence of ABA, while PYL5, PYL6 and PYL8 show partial interactions (Figure 2; Supplementary Figure S1). hab1-pyl6-2 5 HAB1 6 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain Pyl6 9 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified direct interaction direct interaction sf1 mint all receptors tested form stable complexes with DNHAB1 in the presence of ABA at 1:1 stoichiometry, as indicated by the apparent Mr of the complexes (right panels in Figure 2; Supplementary Figure S1). Importantly, PYR1 and PYL1 do not interact with PP2C in the absence of ABA, while PYL5, PYL6 and PYL8 show partial interactions (Figure 2; Supplementary Figure S1). hab1-pyl5-4 5 HAB1 6 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain Pyl5 7 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified direct interaction direct interaction f2 mint all receptors tested form stable complexes with DNHAB1 in the presence of ABA at 1:1 stoichiometry, as indicated by the apparent Mr of the complexes (right panels in Figure 2; Supplementary Figure S1). Importantly, PYR1 and PYL1 do not interact with PP2C in the absence of ABA, while PYL5, PYL6 and PYL8 show partial interactions (Figure 2; Supplementary Figure S1). hab1-pyr1-2 2 Pyr1 11 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified HAB1 6 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain direct interaction direct interaction t2 f5 mint The structure of the ternary complex formed by PYR1H60P, ABA and the catalytic core of HAB1 (DNHAB1) was obtained by X-ray crystallography (see Table II for crystallographic data collection and refinement statistics). The electron density around position 60 of PYR1 confirmed the substitution of histidine for proline (see Supplementary Figure S3). The mutant complex is remarkably similar to that formed by wt PYR1 and DNHAB1 (Dupeux et al, 2011). Atomic coordinates and structure factors for the reported crystal structure have been deposited in the Protein Data Bank under accession code 3ZVU. pyr1-4 4 Pyr1 11 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified Stoichiometry: 2.0 direct interaction direct interaction mint t1 pyr1-4 4 Pyr1 11 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified O49686:p.His60Pro mutation increasing interaction mutation increasing mutation increasing interaction mutation increasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain H P Tue Jul 12 10:55:21 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation increasing' Wed May 02 19:10:11 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'his60pro' Tue Jun 30 15:37:01 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.His60Pro' Stoichiometry: 2.0 direct interaction direct interaction f3c mint we measured dimer dissociation rates by ITC with wt and mutant PYR1. For this purpose, concentrated solutions of either protein were injected into the ITC cell filled with buffer. In this way, the sudden dilution of the protein promotes dissociation and the associated heat can be measured. As can be observed (Figure 3C), the thermogram corresponding to PYR1H60P shows a profile typical of protein dissociation, with endothermic peaks whose area decrease as the protein concentration in the ITC cell increases. The calculated dissociation constant is 39 mM. In contrast, wt PYR1 produced much weaker signals that disappeared after a few injections, indicating that limited dissociation occurs only in extremely diluted conditions. pyr1-4 3 Pyr1 11 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified Stoichiometry: 2.0 direct interaction direct interaction sf2a mint NMR experiments in which binding was followed through ABAinduced resonance shifts on a 15N-labelled PYR1 sample were used to obtain an independent estimation of the dissociation constant. The value of the Kd obtained was 97±36 mM (Supplementary Figure S2). Thus, the dimeric receptor PYR1 has an apparent affinity for ABA that is almost two orders of magnitude lower than that of the monomeric receptors. pyr1-hab1-4 1 Pyr1 11 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell fusion protein fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined O49686:p.His60Pro mutation increasing interaction mutation increasing mutation increasing interaction mutation increasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain H P Tue Jul 12 10:10:49 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation increasing' Wed May 02 18:46:58 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'his60pro' Tue Jun 30 18:02:26 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.His60Pro' HAB1 6 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain fusion protein fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association f3a mint we screened yeast cells for PYR1 mutants that enable interaction with HAB1 in the absence of ABA, which led to the identification of a missense mutant in which histidine 60 was replaced by proline (Figure 3A).