Transient kinetics of ligand binding and role of the C-terminus in the dUTPase from equine infectious anemia virus

Research output: Contribution to journalArticle


Transient kinetics of the equine infectious anemia virus deoxyuridine 5'-triphosphate nucleotide hydrolase were characterized by monitoring the fluorescence of the protein. Rate constants for the association and dissociation of substrate and inhibitors were determined and found to be consistent with a one-step mechanism for substrate binding. A C-terminal part of the enzyme presumed to be flexible was removed by limited trypsinolysis. As a result, the activity of the dUTPase was completely quenched, but the rate constants and fluorescent signal of the truncated enzyme were affected only to a minor degree. We conclude that the flexible C-terminus is not a prerequisite for substrate binding, but indispensable for catalysis.


  • Johan Nord
  • Martin Kiefer
  • Hans-Werner Adolph
  • Michael M Zeppezauer
  • Per-Olof Nyman
Research areas and keywords

Subject classification (UKÄ) – MANDATORY

  • Biological Sciences


  • C-terminus, Deoxyuridine 5'-triphosphate nucleotide hydrolase, Deoxyuridine, Equine infectious anemia virus, Ligand binding, Pre-steady-state kinetics, Rate constant
Original languageEnglish
Pages (from-to)312-316
JournalFEBS Letters
Issue number2-3
Publication statusPublished - 2000
Publication categoryResearch

Related research output

Nord, J., 2000, Johan Nord, Department of Biochemistry, Center for Chemistry and Chemical Engineering, P.O. Box 124, S-221 00, Lund, Sweden. 142 p.

Research output: ThesisDoctoral Thesis (compilation)

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