Biochemistry · 2003 · 62 citations · 5 references
Life SciencesBiochemistryMolecular BiologyDna RepairEnzyme MechanismEnzymologyCatalytic MechanismsDna ChemistryEnzyme KineticsNucleic Acid CleavageElectrophile-protein InteractionsElectrostatic InteractionsEnzymatic Catalysis MechanismsEnzyme Kinetic Isotope EffectsNucleophile ReactivityReaction Coordinate DynamicsReaction Coordinate MappingReactive Electrophile Species
The DNA repair enzyme uracil DNA glycosylase has been crystallized with a cationic 1-aza-2'-deoxyribose-containing DNA that mimics the ultimate transition state of the reaction in which the water nucleophile attacks the anomeric center of the oxacarbenium ion-uracil anion reaction intermediate. Comparison with substrate and product structures, and the previous structure of the intermediate determined by kinetic isotope effects, reveals an exquisite example of geometric strain, least atomic motion, and electrophile migration in biological catalysis. This structure provides a rare opportunity to reconstruct the detailed structural transformations that occur along an enzymatic reaction coordinate.
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