Structural constraints on the ternary complex of 5-enolpyruvylshikimate-3-phosphate synthase from rotational-echo double-resonance NMR.
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.
McDowell
L M
LM
Department of Chemistry Washington University, St. Louis, MO 63130, USA.
Schmidt
A
A
Cohen
E R
ER
Studelska
D R
DR
Schaefer
J
J
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
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