Goal: Understand the chemistry of phosphoester hydrolysis catalyzed by EcoRV and nucleotide ransfer reaction catalyzed by DNA polymerase.
Summary:
Enzymes that make or break phophoester bonds in DNA use metal ions (Mg2+) to activate the nucleophile and stabilize the transition state. Both EcoRV and DNA polymerase hold these Mg2+ ions in place with negatively charged carboxylates of aspartate residues.