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 The crystal structure of galactitol-1-phosphate 5-dehydrogenase from Escherichia coli K12 provides insights into its anomalous behavior on IMAC processes. The crystal structure of galactitol-1-phosphate 5-dehydrogenase from Escherichia coli K12 provides insights into its anomalous behavior on IMAC processes. FEBS Lett. (0014-5793) 2012 Esteban-Torres M., Alvarez Y., Acebrón I., de las Rivas B., Muñoz R., Kohring GW., Roa AM., Sobrino M., Mancheño JM. in vitro In vitro x-ray diffraction x-ray crystallography X-ray X-ray x-ray diffraction predetermined predetermined participant predetermined Esteban-Torres M., Alvarez Y., Acebrón I., de las Rivas B., Muñoz R., Kohring GW., Roa AM., Sobrino M., Mancheño JM. FEBS Lett. (0014-5793) 2012 The crystal structure of galactitol-1-phosphate 5-dehydrogenase from Escherichia coli K12 provides insights into its anomalous behavior on IMAC processes. The crystal structure of galactitol-1-phosphate 5-dehydrogenase from Escherichia coli K12 provides insights into its anomalous behavior on IMAC processes. FEBS Lett. (0014-5793) 2012 Esteban-Torres M., Alvarez Y., Acebrón I., de las Rivas B., Muñoz R., Kohring GW., Roa AM., Sobrino M., Mancheño JM. in vitro In vitro solution sedimentati cosedimentation in solution solution sedimentati predetermined predetermined participant predetermined Esteban-Torres M., Alvarez Y., Acebrón I., de las Rivas B., Muñoz R., Kohring GW., Roa AM., Sobrino M., Mancheño JM. FEBS Lett. (0014-5793) 2012 The crystal structure of galactitol-1-phosphate 5-dehydrogenase from Escherichia coli K12 provides insights into its anomalous behavior on IMAC processes. The crystal structure of galactitol-1-phosphate 5-dehydrogenase from Escherichia coli K12 provides insights into its anomalous behavior on IMAC processes. FEBS Lett. (0014-5793) 2012 Esteban-Torres M., Alvarez Y., Acebrón I., de las Rivas B., Muñoz R., Kohring GW., Roa AM., Sobrino M., Mancheño JM. in vitro In vitro molecular sieving molecular sieving Gel Filtration Size Exclusion Chromatography Size Exclusion Chromatography Gel Filtration Sizing column predetermined predetermined participant predetermined Esteban-Torres M., Alvarez Y., Acebrón I., de las Rivas B., Muñoz R., Kohring GW., Roa AM., Sobrino M., Mancheño JM. FEBS Lett. (0014-5793) 2012 gatd_ecoli Galactitol 1-phosphate 5-dehydrogenase gatD b2091 JW2075 protein protein ecoli Escherichia coli (strain K12) MKSVVNDTDGIVRVAESVIPEIKHQDEVRVKIASSGLCGSDLPRIFKNGAHYYPITLGHEFSGYIDAVGSGVDDLHPGDAVACVPLLPCFTCPECLKGFYSQCAKYDFIGSRRDGGFAEYIVVKRKNVFALPTDMPIEDGAFIEPITVGLHAFHLAQGCENKNVIIIGAGTIGLLAIQCAVALGAKSVTAIDISSEKLALAKSFGAMQTFNSSEMSAPQMQSVLRELRFNQLILETAGVPQTVELAVEIAGPHAQLALVGTLHQDLHLTSATFGKILRKELTVIGSWMNYSSPWPGQEWETASRLLTERKLSLEPLIAHRGSFESFAQAVRDIARNAMPGKVLLIP DIP protein P0A9S3 original sequence version: 39 B35A2B004E5A4032 gatd-1 3 GPDH 4 predetermined predetermined participant predetermined predetermined predetermined participant predetermined unspecified role unspecified role neutral component neutral component purified purified his tag his tag his tag Hexa-His-tag 6-His-tag Histidine-tag Hexa-His-tag Histidine-tag 6-His-tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Stoichiometry: 2.0 direct interaction direct interaction f3a mint Two recombinant variants of the dehydrogenase were produced in E. coli BL21 (DE3) cells: His6-GPDH, which contained an N-terminal His6-tag followed by a TEV cleavage site, and recombinant GPDH, which is equivalent to the endogenous enzyme. The first approach revealed that both variants behave as dimeric species (rGPDH: 63 kDa; His6-GPDH: 72 kDa) and also that His6-GPDH is metastable in solution, forming large aggregates (Fig 3a). gatd-1 2 GPDH 4 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 f3b f3c Conversely, sedimentation velocity analyses revealed that rGPDH (19 and 30 M) is very homogeneous with an s value of 4.5 ± 0.1 S (Fig. 3b). Additionally, the results obtained from the sedimentation equilibrium experiments indicate that the molecular weight of this species (70.7 kDa for the analysis at 14,000 rpm and 77.2 kDa at 18,000 rpm) is consistent with the mass of a dimer (rGPDH monomeric theoretical mass is 37,390 Da; Fig. 3c). This conclusion is definitively supported by the crystal structure of rGPDH which reveals it is a dimeric assembly gatd-1 1 GPDH 4 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 PDB ID: 4A2C t1 f4 sf1