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/ Glutathione degradation by the alternative pathway (DUG pathway) in Saccharomyces cerevisiae is initiated by (Dug2p-Dug3p)2 complex, a novel glutamine amidotransferase (GATase) enzyme acting on glutathione. Glutathione degradation by the alternative pathway (DUG pathway) in Saccharomyces cerevisiae is initiated by (Dug2p-Dug3p)2 complex, a novel glutamine amidotransferase (GATase) enzyme acting on glutathione. The Journal of biological chemistry 2012 imex curation Kaur H., Ganguli D., Bachhawat AK. anand@iisermohali.ac.in 05-Sep-2017: Contacted by IntAct-Help. Only protein-protein interactions imex curation yeasx Saccharomyces cerevisiae (Baker's yeast) 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 Kaur H., Ganguli D., Bachhawat AK. anand@iisermohali.ac.in Accepted 2017-AUG-07 AT 10:22 BST AT 10:22 BST by MGT The Journal of biological chemistry 2012 Only protein-protein interactions imex curation imex curation Amended as per results section Glutathione degradation by the alternative pathway (DUG pathway) in Saccharomyces cerevisiae is initiated by (Dug2p-Dug3p)2 complex, a novel glutamine amidotransferase (GATase) enzyme acting on glutathione. Glutathione degradation by the alternative pathway (DUG pathway) in Saccharomyces cerevisiae is initiated by (Dug2p-Dug3p)2 complex, a novel glutamine amidotransferase (GATase) enzyme acting on glutathione. The Journal of biological chemistry 2012 imex curation Kaur H., Ganguli D., Bachhawat AK. anand@iisermohali.ac.in 05-Sep-2017: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro cleavage cleavage assay cleavage predetermined predetermined participant predetermined Kaur H., Ganguli D., Bachhawat AK. anand@iisermohali.ac.in Accepted 2017-AUG-07 AT 10:22 BST AT 10:22 BST by MGT The Journal of biological chemistry 2012 Only protein-protein interactions imex curation imex curation Amended as per results section Mock publication and experiment for abstract interactions that are not interaction evidences. Mock publication and experiment for abstract interactions that are not interaction evidences. unspecified method Glutathione degradation by the alternative pathway (DUG pathway) in Saccharomyces cerevisiae is initiated by (Dug2p-Dug3p)2 complex, a novel glutamine amidotransferase (GATase) enzyme acting on glutathione. Glutathione degradation by the alternative pathway (DUG pathway) in Saccharomyces cerevisiae is initiated by (Dug2p-Dug3p)2 complex, a novel glutamine amidotransferase (GATase) enzyme acting on glutathione. The Journal of biological chemistry 2012 imex curation Kaur H., Ganguli D., Bachhawat AK. anand@iisermohali.ac.in 05-Sep-2017: Contacted by IntAct-Help. Only protein-protein interactions imex curation ecoli Escherichia coli (strain K12) molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column predetermined predetermined participant predetermined Kaur H., Ganguli D., Bachhawat AK. anand@iisermohali.ac.in Accepted 2017-AUG-07 AT 10:22 BST AT 10:22 BST by MGT The Journal of biological chemistry 2012 Only protein-protein interactions imex curation imex curation Amended as per results section Glutathione degradation by the alternative pathway (DUG pathway) in Saccharomyces cerevisiae is initiated by (Dug2p-Dug3p)2 complex, a novel glutamine amidotransferase (GATase) enzyme acting on glutathione. Glutathione degradation by the alternative pathway (DUG pathway) in Saccharomyces cerevisiae is initiated by (Dug2p-Dug3p)2 complex, a novel glutamine amidotransferase (GATase) enzyme acting on glutathione. The Journal of biological chemistry 2012 imex curation Kaur H., Ganguli D., Bachhawat AK. anand@iisermohali.ac.in 05-Sep-2017: Contacted by IntAct-Help. Only protein-protein interactions imex curation in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column predetermined predetermined participant predetermined Kaur H., Ganguli D., Bachhawat AK. anand@iisermohali.ac.in Accepted 2017-AUG-07 AT 10:22 BST AT 10:22 BST by MGT The Journal of biological chemistry 2012 Only protein-protein interactions imex curation imex curation Amended as per results section dug3_yeast Probable glutamine amidotransferase DUG3 YNL191W N1410 DUG3 GSH degradosomal complex subunit DUG3 Deficient in utilization of glutathione protein 3 protein protein yeast Saccharomyces cerevisiae Baker's yeast MCRFLIFKGKQPIRLSHLLTRPAHSIINQSFDSRLRLDRRRPMNGDGFGVAYYPLDTELSEDGPCLFKAITPAWNNQNLSTLAEKTKSDLVFAHVRASTYGVLSETNCHPFTYHSLCFMHNGGISNFKGIKRKLLNHIKDEYLNFIQGSTDSECAFALFLDTLDKLGYDPKKQDGDFGNVALRKAMLRTIDYIRDWTKEANKDEAHVEPSLLNFAVTDGSTVVVSRYITSKTDEAASLHFSCGSSFVETSPGEYRVERLDRNQDVIMVASEPLTFERGDWTAVPTNSILTIKKQTILLHPIIDEYYQEDPLYLRSSTLAESKGLMGSIPLAKAVEKNVPPLEREGRTRPPTAVAHIA AFAAEFE7152CFE1A AFAAEFE7152CFE1A dug2_yeast Probable di- and tripeptidase DUG2 YBR281C YBR2018 DUG2 GSH degradosomal complex subunit DUG2 Deficient in utilization of glutathione protein 2 protein protein yeast Saccharomyces cerevisiae Baker's yeast MYDSRGVALHSELIHRWNHAFSILSIVAFPKKRLLFAGSQDSKILVFDLPTYNLIHTIRLGESQEETHTRSSVLCLTGSEDENFLFSGGADSLVRIWSIGEKTIRDDFLPVTEIATVYSVTDIGDIFSLAYLDSLETIVFGCQNASLLYVENLIQKIEKKSSDGVENINKLPHRRYDKFFDSLGPTGYSSNSLSQTSLTSLQENCGAAIIEVPSENIIKYAHYGFIYSINKLCPRFNQLLEKSSRTSGAEHIISSAGDGISKLWEFSKDKGQNTVKISLINDKIDNEDSVISQTIEFPFLYCGLTDGIIKIWDLNTQQIISTLKTKHESDVISISVYMDHVFAIDESGITHFYQNQVNHWNPQQGKILSSEIFSKSNAGSVSLLTGGSDGSLTLWDITSLLSAVPLSSNSPINASSTLQTTNLWAAYQSASLNNEEMLNTLRELISFQTVSQSKDTTNTLSLRRCAIYLQQLFLKFGATNSQLFPLPDGGNPVVFAYFQGNGKVSQVKGAKKKRILWYGHYDVISSGNTFNWNTDPFTLTCENGYLKGRGVSDNKGPLVSAIHSVAYLFQQGELVNDVVFLVEGSEEIGSASLKQVCEKYHDIIGKDIDWILLSNSTWVDQEHPCLNYGLRGVINAQIKVWSDKPDGHSGLNGGVYDEPMVNLVKIVSKLQNEQNEIMIPNFYSPLKDLTEEEYQRFQKITELANIDENTTVQDLITNWTKPSLSMTTVKFSGPGNITVIPKSVTMGISIRLVPEQSVEQVKRDLKAYLEESFKQLKSQNHLEIKVLNEAEGWLGDPTNHAYQILKDEITTAWDVEPLLVREGGSISCLRMLERIFDAPAVQIPCGQSTDNGHLANENLRIKNWSNLTEILSKVFNRL A24C10BB4E94C344 A24C10BB4E94C344 l-gamma-glutamyl-l-cysteine L-gamma-glutamyl-L-cysteine 5-L-Glutamyl-L-cysteine gamma-Glu-Cys gamma-Glutamylcysteine gamma-L-Glutamyl-L-cysteine gammaGluCys Glu(-Cys) L-gamma-glutamyl-L-cysteine L-gamma-Glutamylcysteine L-gamma-glutamyl-L-cysteine small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) dug1_yeast Cys-Gly metallodipeptidase DUG1 DUG1 GSH degradosomal complex subunit DUG1 Deficient in utilization of glutathione protein 1 YFR044C protein protein yeast Saccharomyces cerevisiae Baker's yeast MSHSLTSVFQKIDSLKPQFFSRLTKAIQIPAVSSDESLRSKVFDKAKFISEQLSQSGFHDIKMVDLGIQPPPISTPNLSLPPVILSRFGSDPSKKTVLVYGHYDVQPAQLEDGWDTEPFKLVIDEAKGIMKGRGVTDDTGPLLSWINVVDAFKASGQEFPVNLVTCFEGMEESGSLKLDELIKKEANGYFKGVDAVCISDNYWLGTKKPVLTYGLRGCNYYQTIIEGPSADLHSGIFGGVVAEPMIDLMQVLGSLVDSKGKILIDGIDEMVAPLTEKEKALYKDIEFSVEELNAATGSKTSLYDKKEDILMHRWRYPSLSIHGVEGAFSAQGAKTVIPAKVFGKFSIRTVPDMDSEKLTSLVQKHCDAKFKSLNSPNKCRTELIHDGAYWVSDPFNAQFTAAKKATKLVYGVDPDFTREGGSIPITLTFQDALNTSVLLLPMGRGDDGAHSINEKLDISNFVGGMKTMAAYLQYYSESPEN 3E53773A945F5EBC 3E53773A945F5EBC glutathione glutathione gamma-L-Glutamyl-L-cysteinyl-glycine 5-L-Glutamyl-L-cysteinylglycine Glutathione Glutathione-SH GSH N-(N-gamma-L-Glutamyl-L-cysteinyl)glycine Reduced glutathione L-gamma-glutamyl-L-cysteinylglycine small molecule small molecule chemical synthesis Chemical synthesis (Chemical synthesis) dug2-dug3-1 1 6 unspecified role unspecified role bait bait 7 unspecified role unspecified role prey prey wd region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain physical association physical association Supp. Fig. 1 dug2-dug2-1 1 7 unspecified role unspecified role prey prey 7 unspecified role unspecified role bait bait physical association physical association Supp. Fig. 1 dug2-1 4 7 unspecified role unspecified role neutral component neutral component n-terminal 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 m20a region sufficient to bind sufficient binding region sufficient to bind certain certain sequence position certain certain certain sequence position certain direct interaction direct interaction Fig. 1A, 1B, Supp. Fig. 2 dug1-1 4 9 unspecified role unspecified role neutral component neutral component direct interaction direct interaction Supp. Fig. 2 dug2-dug3-2 5 7 unspecified role unspecified role neutral component neutral component n-terminal 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 unspecified role unspecified role neutral component neutral component c-terminal his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag c-term range C-terminal range c-term range c-term range C-terminal range c-term range P53871:p.Ala357delinsAlaGlyGlyGlyGlyGlyGlyIlePro mutation mutation certain certain sequence position certain certain certain sequence position certain A AGGGGGGIP Wed May 02 15:59:03 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ala357ala_gly_gly_gly_gly_gly_gly_ile_pro' Wed Jun 13 17:48:40 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ala357delinsAlaGlyGlyGlyGlyGlyGlyIlePro' Tue Jun 30 19:09:41 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ala357delinsAlaGlyGlyGlyGlyGlyGlyIlePro' ecoli Escherichia coli (strain K12) direct interaction direct interaction Fig. 5A dug2_dug3_yeast-glutathione-1 2 10 enzyme target enzyme target substrate substrate unspecified role unspecified role 33 enzyme enzyme unspecified role unspecified role yeast Saccharomyces cerevisiae Baker's yeast cleavage cleavage reaction cleavage Fig. 4, 5B dug2_dug3_yeast-glutathione-2 2 8 enzyme target enzyme target substrate substrate unspecified role unspecified role chemical synthesis Chemical synthesis (Chemical synthesis) 33 enzyme enzyme unspecified role unspecified role yeast Saccharomyces cerevisiae Baker's yeast cleavage cleavage reaction cleavage Supp Fig. 4 dug2_dug3_yeast Glutathione hydrolase complex DUG2:DUG3 GSH degradosomal complex 7 unspecified role unspecified role dug2 homodimerisation region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain true dug3_binding_region dug3 binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain 7 unspecified role unspecified role dug2 homodimerisation region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain true dug3_binding_region dug3 binding region binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain 6 enzyme enzyme GATase glutamine amidotransferase biological feature biological feature certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain dug2_binding_region dug2 binding region binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 6 unspecified role unspecified role GATase glutamine amidotransferase biological feature biological feature certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain certain certain sequence position certain dug2_binding_region dug2 binding region binding region binding-associated region binding region undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined 41 38 39 48 45 42 physical association physical association yeast Saccharomyces cerevisiae Baker's yeast stable complex stable complex physical interaction evidence used in manual assertion physical interaction evidence used in manual assertion IPI inferred from physical interaction Glutathione hydrolase/Glutamine Amidotransferase II complex responsible for glutathione degradation. Both DUG2 and DUG3 are depressed under sulfur limitation conditions. Although neither protein alone can cleave glutathione, the complex degrades glutathione by cleaving of the gamma-glutamyl linkage. DUG3 has the GATase activity, while DUG2 probably acts as a scaffold protein, bringing two DUG3 proteins together. Purified complex is approximately 300 kDa with a Km of 1.2mM. Accepted 2016-JAN-07 by ORCHARD