Specific derivatization of the active site tyrosine in dUTPase perturbs ligand binding to the active site

B. Vértessy, Rebecca Persson, Anna Maria Rosengren, Michael Zeppezauer, Per Olof Nyman

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

Selective modification of one (of three) tyrosine residue per enzyme monomer lends to inactivation of dUTPase of the retrovirus equine infectious anemia virus (EIAV). The substrate dUMP and the cofactor Mg2+ protects against inactivation and modification, in agreement with the study on E. coli dUTPase. Amino acid analyses of nitrated dUTPases confirmed Tyr-selectivity of modification. The nitrated residue in E. coli dUTPase was identified as the evolutionary highly conserved Tyr-93. The modifiable residue is shown to be the only Tyr exposed in both E. coli and EIAV dUTPases. As a consequence of Tyr-93 derivatization, the Mg2+-dependent interaction between the substrate-analogue dUDP and E. coli dUTPase becomes impaired as shown by circular dichroism spectroscopy, here presented as a tool for monitoring ligand binding to the active site.

Original languageEnglish
Pages (from-to)294-300
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume219
Issue number2
DOIs
Publication statusPublished - Feb 15 1996

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Escherichia coli
Tyrosine
Catalytic Domain
Equine infectious anemia virus
Ligands
Viruses
Circular dichroism spectroscopy
Substrates
Retroviridae
Circular Dichroism
Spectrum Analysis
Monomers
Amino Acids
dUTP pyrophosphatase
Monitoring
Enzymes

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Specific derivatization of the active site tyrosine in dUTPase perturbs ligand binding to the active site. / Vértessy, B.; Persson, Rebecca; Rosengren, Anna Maria; Zeppezauer, Michael; Nyman, Per Olof.

In: Biochemical and Biophysical Research Communications, Vol. 219, No. 2, 15.02.1996, p. 294-300.

Research output: Contribution to journalArticle

Vértessy, B. ; Persson, Rebecca ; Rosengren, Anna Maria ; Zeppezauer, Michael ; Nyman, Per Olof. / Specific derivatization of the active site tyrosine in dUTPase perturbs ligand binding to the active site. In: Biochemical and Biophysical Research Communications. 1996 ; Vol. 219, No. 2. pp. 294-300.
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