Enhanced expression of the Escherichia coli serA gene in a plasmid vector. Purification, crystallization, and preliminary X-ray data of D-3 phosphoglycerate dehydrogenase.
The serA gene of Escherichia coli strain K-12, which codes for the cooperative allosteric enzyme D-3-phosphoglycerate dehydrogenase, was inserted into an inducible expression vector which produced phosphoglycerate dehydrogenase as 8% of the soluble protein of E. coli. The purified protein was used to grow several different single crystal forms. One of these, with space group P2(1), appears to contain all four subunits of the tetrameric enzyme in the asymmetric unit and diffracts to sufficient resolution to allow determination of the structure of phosphoglycerate dehydrogenase.
Schuller
D J
DJ
Department of Biological Chemistry, Washington University School of Medicine, St. Louis, Missouri 63110.
Fetter
C H
CH
Banaszak
L J
LJ
Grant
G A
GA
D002237:Carbohydrate Dehydrogenases / Q000235:genetics* / Q000302:isolation & purification; D003460:Crystallization; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D005796:Genes; D005798:Genes, Bacterial; D005822:Genetic Vectors; D046911:Macromolecular Substances; D050543:Phosphoglycerate Dehydrogenase; D010957:Plasmids; D015183:Restriction Mapping; D014961:X-Ray Diffraction
D046911:Macromolecular Substances; D002237:Carbohydrate Dehydrogenases; D050543:Phosphoglycerate Dehydrogenase