Time-resolved solid-state REDOR NMR studies of UDP N-acetylglucosamine enolpyruvyl transferase.
The new method of time-resolved solid-state rotational echo double resonance (REDOR) NMR spectroscopy introduced recently by this laboratory has been applied to the enzyme uridine N-acetylglucosamine (UDP-NAG) enolpyruvyl transferase (EPT), with the goal of probing the interactions between reactive species and their enzyme active site. The approach has been used in a qualitative fashion with the enzyme-inhibitor and enzyme-intermediate complexes of uniformly 15N-labeled UDP-NAG EPT, trapped under steady-state and pre-steady-state conditions. A different set of intermolecular interactions between the substrates UDP-NAG, UDP-NAG plus 3-Z-fluorophosphoenolpyruvate, covalent O-phosphothioketal, and UDP-NAG plus phosphoenolpyruvate trapped under time-resolved conditions (after 50 ms reaction time), and the EPT enzyme active site were observed, and this is contrasted to a similar study of the interactions in a related enzyme, 5-enolpyruvyl-shikimate-3-phosphate synthase.
Department of Biochemistry and Biophysics, Washington State University, Pullman 99164-4660, USA.
Y
Y
Li
F
F
Krekel
C A
CA
Ramilo
N
N
Amrhein
J N
JN
Evans
D019883:Alkyl and Aryl Transferases; D001665:Binding Sites; D009682:Magnetic Resonance Spectroscopy; D015394:Molecular Structure; D013379:Substrate Specificity; D014166:Transferases / Q000737:chemistry* / Q000378:metabolism; D014537:Uridine Diphosphate N-Acetylglucosamine / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism
D014537:Uridine Diphosphate N-Acetylglucosamine; C038277:UDP-N-acetylglucosamine-enolpyruvate; D014166:Transferases; D019883:Alkyl and Aryl Transferases; C022566:UDP-N-acetylglucosamine 1-carboxyvinyltransferase