Active site closure facilitates juxtaposition of reactant atoms for initiation of catalysis by human dUTPase

Balázs Varga, Orsolya Barabás, Júlia Kovári, J. Tóth, E. Hunyadi-Gulyás, E. Klement, Katalin F. Medzihradszky, F. Tölgyesi, J. Fidy, B. Vértessy

Research output: Contribution to journalArticle

46 Citations (Scopus)

Abstract

Human dUTPase, essential for DNA integrity, is an important survival factor for cancer cells. We determined the crystal structure of the enzyme:α,β-imino-dUTP:Mg complex and performed equilibrium binding experiments in solution. Ordering of the C-terminus upon the active site induces close juxtaposition of the incoming nucleophile attacker water oxygen and the α-phosphorus of the substrate, decreasing their distance below the van der Waals limit. Complex interactions of the C-terminus with both substrate and product were observed via a specifically designed tryptophan sensor, suitable for further detailed kinetic and ligand binding studies. Results explain the key functional role of the C-terminus.

Original languageEnglish
Pages (from-to)4783-4788
Number of pages6
JournalFEBS Letters
Volume581
Issue number24
DOIs
Publication statusPublished - Oct 2 2007

Fingerprint

Catalysis
Tryptophan
Phosphorus
Catalytic Domain
Oxygen
Ligands
Atoms
Nucleophiles
Survival
Water
DNA
Substrates
Enzymes
Neoplasms
Crystal structure
Cells
Kinetics
Sensors
Experiments
dUTP pyrophosphatase

Keywords

  • Enzyme mechanism
  • Flexible C-terminus
  • High-throughput screening for ligand binding
  • Initiation of catalysis
  • Tryptophan sensor

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Active site closure facilitates juxtaposition of reactant atoms for initiation of catalysis by human dUTPase. / Varga, Balázs; Barabás, Orsolya; Kovári, Júlia; Tóth, J.; Hunyadi-Gulyás, E.; Klement, E.; Medzihradszky, Katalin F.; Tölgyesi, F.; Fidy, J.; Vértessy, B.

In: FEBS Letters, Vol. 581, No. 24, 02.10.2007, p. 4783-4788.

Research output: Contribution to journalArticle

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AU - Tóth, J.

AU - Hunyadi-Gulyás, E.

AU - Klement, E.

AU - Medzihradszky, Katalin F.

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