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Updated: Jun 11, 2026
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Nucleoside Triphosphates - From Synthesis to Biochemical Characterization
Published on: April 3, 2014
Aromatic stacking between nucleobase and enzyme promotes phosphate ester hydrolysis in dUTPase
Ildiko Pecsi1, Ibolya Leveles, Veronika Harmat
1Institute of Enzymology, Biological Research Center, Hungarian Academy of Sciences, Budapest, Hungary.
Nucleic Acids Research
|July 6, 2010
Summary
A conserved aromatic stacking interaction in dUTPase stabilizes the transition state for nucleotide hydrolysis. This remote π-π interaction accelerates catalysis, potentially in many nucleotide-hydrolyzing enzymes.
Area of Science:
- Biochemistry
- Enzymology
- Molecular Recognition
Background:
- Aromatic interactions are crucial for molecular recognition but their catalytic roles in biology are not well-understood.
- The specific contribution of aromatic stacking to enzyme catalysis, particularly in nucleotide hydrolysis, remains largely undocumented.
Purpose of the Study:
- To investigate the role of a conserved aromatic stacking interaction between dUTPase and its nucleotide substrate in catalysis.
- To elucidate the mechanism by which this remote aromatic interaction influences the nucleotide hydrolysis reaction and accelerates its rate.
Main Methods:
- Crystallography
- Enzyme kinetics assays
- Optical spectroscopy
- Thermodynamic calculations
Main Results:
- A conserved aromatic stacking interaction significantly stabilizes the associative transition state of nucleotide hydrolysis in dUTPase.
- This π-π interaction accelerates catalysis despite the aromatic moiety (uracil) being distant from the hydrolysis site (α-phosphate).
Conclusions:
- A remote π-π interaction mechanism contributes to enzymatic rate acceleration in nucleotide hydrolysis.
- This phenomenon may be a general feature in the catalysis of phosphate ester hydrolysis by various nucleotide-hydrolyzing enzymes, including ATPases.
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