title: Analysis of fluoromethyl group chirality establishes a common stereochemical course for the enolpyruvyl transfers catalyzed by EPSP synthase and UDP-GlcNAc enolpyruvyl transferase.
abstract: The stereochemistry of transient methyl group formation at C-3 of phosphoenolpyruvate (PEP) in the reaction catalyzed by 5-enolpyruvylshikimate 3-phosphate (EPSP) synthase has been examined using the pseudosubstrates, (E)- and (Z)-3-fluorophosphoenolpyruvate (FPEP). Kinetically stable, chiral [1H, 2H]fluoromethyl analogs of the reaction tetrahedral intermediate were isolated and subjected to decomposition and stereochemical analysis. EPSP synthase was found to catalyze the 2-re face addition of solvent-derived hydrogen to C-3 of FPEP (corresponding to the 2-si face of PEP). Comparison of these data with prior analogous work on the MurA reaction [Kim, D.H., Lees, W.J., & Walsh, C. T. (1995) J. Am. Chem. Soc. 117, 6380-6381] suggests that the two enolpyruvyl transferases share a common stereochemical course, further strengthening the mechanistic, structural, and evolutionary relationship between the two enzymes.
authors.affiliation: Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, Massachusetts 02115, USA.
authors.forename: D H
authors.initials: DH
authors.lastname: Kim
authors.forename: G W
authors.initials: GW
authors.lastname: Tucker-Kellogg
authors.forename: W J
authors.initials: WJ
authors.lastname: Lees
authors.forename: C T
authors.initials: CT
authors.lastname: Walsh
mesh_terms: D051229:3-Phosphoshikimate 1-Carboxyvinyltransferase; D019883:Alkyl and Aryl Transferases; D001483:Base Sequence; D017931:DNA Primers / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D010728:Phosphoenolpyruvate / Q000378:metabolism; D011994:Recombinant Proteins; D012765:Shikimic Acid / Q000031:analogs & derivatives / Q000378:metabolism; D013237:Stereoisomerism; D014166:Transferases / Q000737:chemistry*
chemical_list: D017931:DNA Primers; D011994:Recombinant Proteins; D012765:Shikimic Acid; C042786:shikimic acid-3-phosphate; D010728:Phosphoenolpyruvate; D014166:Transferases; D019883:Alkyl and Aryl Transferases; D051229:3-Phosphoshikimate 1-Carboxyvinyltransferase; C022566:UDP-N-acetylglucosamine 1-carboxyvinyltransferase