title: 3-Phosphoglycerate dehydrogenase from Corynebacterium glutamicum: the C-terminal domain is not essential for activity but is required for inhibition by L-serine.
abstract: The serA gene of Corynebacterium glutamicum coding for 3-phosphoglycerate dehydrogenase (PGDH) was isolated and functionally characterized. It encodes a polypeptide of 530 aminoacyl residues (aa), which is substantially longer than the corresponding Escherichia coli polypeptide of 410 aa. The difference is largely due to an additional stretch of aa in the carboxy- (C)-terminal part of the polypeptide. Overexpression of serA in C. glutamicum results in a 16-fold increase in specific PGDH activity to 2.1 U/mg protein, with activity being inhibited by high concentrations of L-serine. A set of muteins that were progressively truncated at the C-terminal end was constructed. When overexpressed, mutein SerADelta197 showed a specific PGDH dehydrogenase activity of 1.3 U/mg protein, with the activity no longer being sensitive to L-serine. Gel filtration experiments showed that wild type PGDH is a homotetramer, whereas mutein SerADelta197 constitutes a dimer. Thus, the specific regulatory features of C. glutamicum PGDH are due to the C-terminal part of the polypeptide, which can be deleted with almost no effect on the catalytic activity of the enzyme.
authors.affiliation: Institut für Biotechnologie 1, Forschungszentrum Jülich GmbH, Germany, p.wendisch@fz-juelich.de
authors.forename: P
authors.initials: P
authors.lastname: Peters-Wendisch
authors.forename: R
authors.initials: R
authors.lastname: Netzer
authors.forename: L
authors.initials: L
authors.lastname: Eggeling
authors.forename: H
authors.initials: H
authors.lastname: Sahm
mesh_terms: D002237:Carbohydrate Dehydrogenases / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D003352:Corynebacterium / Q000201:enzymology* / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D050543:Phosphoglycerate Dehydrogenase; D017434:Protein Structure, Tertiary / Q000502:physiology; D012694:Serine / Q000494:pharmacology*; D013329:Structure-Activity Relationship
chemical_list: D012694:Serine; D002237:Carbohydrate Dehydrogenases; D050543:Phosphoglycerate Dehydrogenase