PMID:1939260
| Citation |
Padgette, SR, Re, DB, Gasser, CS, Eichholtz, DA, Frazier, RB, Hironaka, CM, Levine, EB, Shah, DM, Fraley, RT and Kishore, GM (1991) Site-directed mutagenesis of a conserved region of the 5-enolpyruvylshikimate-3-phosphate synthase active site. J. Biol. Chem. 266:22364-9 |
|---|---|
| Abstract |
The active site of the enzyme 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) has been probed using site-directed mutagenesis and inhibitor binding techniques. Replacement of a specific glycyl with an alanyl or a prolyl with a seryl residue in a highly conserved region confers glyphosate tolerance to several bacterial and plant EPSPS enzymes, suggesting a high degree of structural conservation between these enzymes. The glycine to alanine substitution corresponding to Escherichia coli EPSPS G96A increases the Ki(app) (glyphosate) of petunia EPSPS 5000-fold while increasing the Km(app)(phosphoenolpyruvate) about 40-fold. Substitution of this glycine with serine, however, abolishes EPSPS activity but results in the elicitation of a novel EPSP hydrolase activity whereby EPSP is converted to shikimate 3-phosphate and pyruvate. This highly conserved region is critical for the interaction of the phosphate moiety of phosphoenolpyruvate with EPSPS. |
| Links | |
| Keywords |
3-Phosphoshikimate 1-Carboxyvinyltransferase; Alkyl and Aryl Transferases; Amino Acid Sequence; Bacteria/enzymology; Bacteria/genetics; Binding Sites; Biological Evolution; Escherichia coli/enzymology; Escherichia coli/genetics; Kinetics; Molecular Sequence Data; Mutagenesis, Site-Directed; Plants/enzymology; Plants/genetics; Plasmids; Saccharomyces cerevisiae/enzymology; Saccharomyces cerevisiae/genetics; Sequence Homology, Nucleic Acid; Species Specificity; Transferases/genetics; Transferases/isolation & purification; Transferases/metabolism |
| edit table |
Significance
You can help EcoliWiki by summarizing why this paper is useful
Useful Materials and Methods
You can help Ecoliwiki by describing the useful materials (strains, plasmids, antibodies, etc) described in this paper.
Annotations
<annotationlinks/>
EcoliWiki Links
Add links to pages that link here (e.g. gene, product, method pages)
References
See Help:References for how to manage references in EcoliWiki.