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Time‐resolved solid‐state REDOR NMR studies of UDP N ‐acetylglucosamine enolpyruvyl transferase
Author(s) -
Li Yan,
Krekel Florian,
Ramilo Cecilia A.,
Amrhein Nikolaus,
Evans Jeremy N.S.
Publication year - 1995
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(95)01338-5
Subject(s) - enzyme , transferase , chemistry , active site , biosynthesis , atp synthase , phosphoenolpyruvate carboxykinase , nuclear magnetic resonance spectroscopy , stereochemistry , biochemistry
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 15 N‐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.