title: Structural constraints on the ternary complex of 5-enolpyruvylshikimate-3-phosphate synthase from rotational-echo double-resonance NMR.
abstract: The 46 kDa enzyme 5-enolpyruvylshikimate-3-phosphate (EPSP) synthase catalyzes the condensation of shikimate-3-phosphate (S3P) and phosphoenolpyruvate to form EPSP. The reaction is inhibited by N-(phosphonomethyl)-glycine (Glp), which in the presence of S3P, binds to EPSP synthase to form a stable ternary complex. As part of a solid-state NMR characterization of this structure, 15N labels were introduced selectively into the lysine, arginine and histidine residues of EPSP synthase and distances to a 13C label in Glp and to the 31P in S3P and Glp were measured by rotational-echo double-resonance NMR. Three lysine and four arginine residues are in the proximity of the phosphate group of S3P and the carboxyl and phosphonate groups of Glp. A single histidine residue is in the vicinity of the binding site (closer to Glp than to S3P) but is more distant than the lysine and arginine residues.
authors.affiliation: Department of Chemistry Washington University, St. Louis, MO 63130, USA.
authors.forename: L M
authors.initials: LM
authors.lastname: McDowell
authors.forename: A
authors.initials: A
authors.lastname: Schmidt
authors.forename: E R
authors.initials: ER
authors.lastname: Cohen
authors.forename: D R
authors.initials: DR
authors.lastname: Studelska
authors.forename: J
authors.initials: J
authors.lastname: Schaefer
mesh_terms: D051229:3-Phosphoshikimate 1-Carboxyvinyltransferase; D019883:Alkyl and Aryl Transferases; D001120:Arginine / Q000737:chemistry; D001665:Binding Sites; D004926:Escherichia coli / Q000201:enzymology; D005998:Glycine / Q000031:analogs & derivatives / Q000378:metabolism; D006639:Histidine / Q000737:chemistry; D008239:Lysine / Q000737:chemistry; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D015394:Molecular Structure; D009587:Nitrogen Isotopes; D012765:Shikimic Acid / Q000031:analogs & derivatives / Q000378:metabolism; D014166:Transferases / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism; D000097797:Glyphosate
chemical_list: D051229:3-Phosphoshikimate 1-Carboxyvinyltransferase; D019883:Alkyl and Aryl Transferases; D001120:Arginine; D005998:Glycine; D006639:Histidine; D008239:Lysine; D009587:Nitrogen Isotopes; D012765:Shikimic Acid; D014166:Transferases; D000097797:Glyphosate; C042786:shikimic acid-3-phosphate