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A spectrophotometric assay for quantitative determination of kcat of herpes simplex virus type 1 thymidine kinase substrates.
Journal article

A spectrophotometric assay for quantitative determination of kcat of herpes simplex virus type 1 thymidine kinase substrates.

  • Schelling P Department of Applied BioSciences, Swiss Federal Institute of Technology (ETH), Winterthurerstrasse 190, Zürich, CH-8057, Switzerland.
  • Folkers G
  • Scapozza L
  • 2001-07-31
Published in:
  • Analytical biochemistry. - 2001
Animals
Antiviral Agents
Catalysis
Drug Design
Herpesvirus 1, Human
Kinetics
L-Lactate Dehydrogenase
NAD
Phosphoenolpyruvate
Phosphorylation
Pyruvate Kinase
Rabbits
Radiometry
Reproducibility of Results
Spectrophotometry, Ultraviolet
Thymidine Kinase
English A simple method to determine the in vitro catalytic turnover constant of several substrates of herpes simplex virus type 1 thymidine kinase is presented in this study. The method is based on a continuous spectroscopic enzyme-coupled assay and allows one to monitor the herpes simplex virus type 1 thymidine kinase activity in the presence of unlabeled substrates. A clear correlation between the catalytic turnover constant and the rate of decrease in absorbance over time during the assay has been demonstrated. Exploiting this correlation, this method has been used to determine rapidly and precisely the catalytic turnover constant of antiviral lead compounds not readily available in the radioactive labeled form.
Language
  • English
Open access status
closed
Identifiers
Persistent URL
https://sonar.ch/global/documents/52510
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