Structural Constraints on the Ternary Complex of 5-Enolpyruvylshikimate-3-phosphate Synthase from Rotational-echo Double-resonance NMR

• Published in: Journal of molecular biology 1996-02, Vol.256 (1), p.160-171
• Main Authors: McDowell, Lynda M., Schmidt, Asher, Cohen, Eric R., Studelska, Daniel R., Schaefer, Jacob
• Format: Article
• Language: English
• Subjects: 3-Phosphoshikimate 1-Carboxyvinyltransferase; 5-enolpyruvylshikimate-3-phosphate synthase; Alkyl and Aryl Transferases; Arginine - chemistry; Binding Sites; Escherichia coli - enzymology; Glycine - analogs & derivatives; Glycine - metabolism; Glyphosate; Histidine - chemistry; interatomic distances; Lysine - chemistry; Magnetic Resonance Spectroscopy - methods; Molecular Structure; Nitrogen Isotopes; protein binding site; Shikimic Acid - analogs & derivatives; Shikimic Acid - metabolism; solid-state NMR; stable-isotope label; Transferases - antagonists & inhibitors; Transferases - chemistry; Transferases - metabolism
• ISSN: 0022-2836; 1089-8638
• DOI: 10.1006/jmbi.1996.0074

The 46 kDa enzyme 5-enolpyruvylshikimate-3-phosphate (EPSP) synthase catalyzes the condensation of shikimate-3-phosphate (S3P) and phospho enolpyruvate to form EPSP. The reaction is inhibited by N-(phosphono- methyl)-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.