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 Glutamine synthetase isoforms in nitrogen-fixing soybean nodules: Distinct oligomeric structures and thiol-based regulation. Glutamine synthetase isoforms in nitrogen-fixing soybean nodules: Distinct oligomeric structures and thiol-based regulation. FEBS Lett. (0014-5793) 2015 imex curation Masalkar P.D., Roberts D.M. drobert2@utk.edu Only protein-protein interactions in vitro In vitro solution sedimentati cosedimentation in solution solution sedimentati predetermined predetermined participant predetermined Masalkar P.D., Roberts D.M. drobert2@utk.edu Accepted 2015-FEB-23 by LUANA FEBS Lett. (0014-5793) 2015 Only protein-protein interactions imex curation Glutamine synthetase isoforms in nitrogen-fixing soybean nodules: Distinct oligomeric structures and thiol-based regulation. Glutamine synthetase isoforms in nitrogen-fixing soybean nodules: Distinct oligomeric structures and thiol-based regulation. FEBS Lett. (0014-5793) 2015 imex curation Masalkar P.D., Roberts D.M. drobert2@utk.edu Only protein-protein interactions in vitro In vitro comig non denat gel comigration in non denaturing gel electrophoresis comig non denat gel weight by staining molecular weight estimation by staining weight by staining Masalkar P.D., Roberts D.M. drobert2@utk.edu Accepted 2015-FEB-23 by LUANA FEBS Lett. (0014-5793) 2015 Only protein-protein interactions imex curation Glutamine synthetase isoforms in nitrogen-fixing soybean nodules: Distinct oligomeric structures and thiol-based regulation. Glutamine synthetase isoforms in nitrogen-fixing soybean nodules: Distinct oligomeric structures and thiol-based regulation. FEBS Lett. (0014-5793) 2015 imex curation Masalkar P.D., Roberts D.M. drobert2@utk.edu Only protein-protein interactions in vitro In vitro bn-page blue native page bn-page weight by comassie molecular weight estimation by coomasie staining weight by comassie Masalkar P.D., Roberts D.M. drobert2@utk.edu Accepted 2015-FEB-23 by LUANA FEBS Lett. (0014-5793) 2015 Only protein-protein interactions imex curation Glutamine synthetase isoforms in nitrogen-fixing soybean nodules: Distinct oligomeric structures and thiol-based regulation. Glutamine synthetase isoforms in nitrogen-fixing soybean nodules: Distinct oligomeric structures and thiol-based regulation. FEBS Lett. (0014-5793) 2015 imex curation Masalkar P.D., Roberts D.M. drobert2@utk.edu 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 Masalkar P.D., Roberts D.M. drobert2@utk.edu Accepted 2015-FEB-23 by LUANA FEBS Lett. (0014-5793) 2015 Only protein-protein interactions imex curation q9fuk4_soybn Glutamine synthetase GLYMA_11G215500 548082 protein protein soybn Glycine max (Soybean) Glycine hispida MSLLSDLINLNLSDTTEKVIAEYIWIGGSGMDLRSKARTLPGPVSDPSELPKWNYDGSSTGQAPGEDSEVILYPQAIFRDPFRRGNNILVICDAYTPAGEPIPTNKRHAAAKVFSHPDVVAEVPWYGIEQEYTLLQKDIQWPLGWPVGGFPGPQGPYYCGVGADKAFGRDIVDAHYKACIYAGINISGINGEVMPGQWEFQVGPSVGISAGDEIWAARYILERITEIAGVVVSFDPKPIKGDWNGAGAHTNYSTKSMREDGGYEVIKAAIDKLGKKHKEHIAAYGEGNERRLTGRHETADINTFLWGVANRGASVRVGRDTEKAGKGYFEDRRPASNMDPYVVTSMIADTTILWKP 14BF92D0786FFBB5 glna2_soybn Glutamine synthetase cytosolic isozyme 2 GS1-2 Glutamate--ammonia ligase protein protein soybn Glycine max (Soybean) Glycine hispida MSLLSDLINLNLSDITDKVIAEYIWVGGSGMDMRSKARTLSGPVKDPSKLPKWNYDGSSTGQAPGQDSEVILYPQAIFKDPFRRGSNILVMCDAYTPAGEPIPTNKRNNAAKIFGHPDVAAEEPWYGLEQEYTLLQKDVQWPLGWPLGGFPGPQGPYYCGTGANKAFGRDIVDSHYKACIYAGINISGINGEVMPGQWEFQVGPSIGISAADELWVARYILERITEIAGVVLSFDPKPIQGDWNGAGAHTNYSTKSMRNDGGYEVIKKAIAKLEKRHKEHIAAYGEGNERRLTGRHETADMNTFVWGVANRGASIRVGRDTEKAGKGYFEDRRPASNMDPYVVTSMIAETTILWKP 4B9C63B7718656B5 q9fuk4 1 5 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role unspecified role unspecified role purified purified direct interaction direct interaction f4b t1 sf3a The molecular weight calculated from equilibrium AUC for GS1c1 is 499.28kDa whereas GS1b1 is 409.55kDa (Table 1), supporting the results of velocity AUC. Considering the monomeric molecular weight of both isoforms, the 499.28kDa GS1c1 would represent an oligomer of 12 subunits, which is distinct from the 409.55kDa GS1b1 which would represent a decameric enzyme similar to maize GS1a enzyme q9fuk4-1 1 6 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role unspecified role unspecified role purified purified direct interaction direct interaction f4b t1 sf3b The molecular weight calculated from equilibrium AUC for GS1c1 is 499.28kDa whereas GS1b1 is 409.55kDa (Table 1), supporting the results of velocity AUC. Considering the monomeric molecular weight of both isoforms, the 499.28kDa GS1c1 would represent an oligomer of 12 subunits, which is distinct from the 409.55kDa GS1b1 which would represent a decameric enzyme similar to maize GS1a enzyme glna2-glna2 2 6 weight by staining molecular weight estimation by staining weight by staining weight by staining molecular weight estimation by staining weight by staining unspecified role unspecified role neutral component neutral component purified purified O82560:p.Cys159Ser mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain C S Wed May 02 15:34:25 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys159ser' Tue Jun 30 17:42:24 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Cys159Ser' 6 weight by staining molecular weight estimation by staining weight by staining weight by staining molecular weight estimation by staining weight by staining unspecified role unspecified role neutral component neutral component purified purified O82560:p.Cys159Ser mutation disrupting mutation disrupting interaction mutation disrupting certain certain sequence position certain certain certain sequence position certain C S Wed May 02 15:58:36 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'cys159ser' Tue Jun 30 17:10:29 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Cys159Ser' direct interaction direct interaction f1b f3d After exposure to oxidizing conditions (air oxidation or H2O2 incubation), purified GS1c1C159S and GS1c1 were analyzed by non-reducing SDS–PAGE to determine the effect of oxidation on oligomerization. In contrast to wild type GS1c1, high molecular weight oligomeric species were not observed in the GS1c1C159S mutant under oxidizing conditions (Fig. 3D). glna2-glna2-1 3 6 weight by comassie molecular weight estimation by coomasie staining weight by comassie weight by comassie molecular weight estimation by coomasie staining weight by comassie unspecified role unspecified role neutral component neutral component purified purified 6 weight by comassie molecular weight estimation by coomasie staining weight by comassie weight by comassie molecular weight estimation by coomasie staining weight by comassie unspecified role unspecified role neutral component neutral component purified purified direct interaction direct interaction f4a Comparison of the two GS1 isoforms by native PAGE showed differences in electrophoretic mobility. GS1c1 has a single band with a reduced electrophoretic mobility compared to the major band in GS1b1 (Fig. 4A). A minor species of GS1b1 with a similar electrophoretic mobility as GS1c1 was also observed, suggesting the presence of a small amount of a second oligomeric species with similar properties to the c isoform. Nevertheless, the results show fundamental differences in the physicochemical properties of the two isoforms. glna2-glna2-2 4 6 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified 6 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified direct interaction direct interaction sf2 The possibility that oligomerization was the result of an intersubunit disulfide between cys92 and cys159, was investigated by site-directed mutagenesis in which each cysteine residue was replaced by serine. Of the two cys-to-ser mutants, only GS1- c1C159S produced an active oligomeric protein (Supplementary Fig. S2).