Fluorescing Isofunctional Ribonucleosides: Assessing Adenosine Deaminase Activity and Inhibition

Chembiochem. 2019 Mar 1;20(5):718-726. doi: 10.1002/cbic.201800665. Epub 2019 Feb 21.

Abstract

The enzymatic conversion of isothiazolo[4,3-d]pyrimidine-based adenosine (tz A) and 2-aminoadenosine (tz 2-AA) analogues to the corresponding isothiazolo[4,3-d]pyrimidine-based inosine (tz I) and guanosine (tz G) derivatives is evaluated and compared to the conversion of native adenosine to inosine. Henri-Michaelis-Menten analyses provides the foundation for a high-throughput screening assay, and the efficacy of the assay is showcased by fluorescence-based analysis of tz A conversion to tz I in the presence of known and newly synthesized inhibitors.

Keywords: adenosine; adenosine deaminase; enzyme catalysis; high-throughput screening; inhibitors.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Adenosine Deaminase / metabolism*
  • Adenosine* / analogs & derivatives
  • Adenosine* / metabolism
  • Enzyme Inhibitors / chemistry
  • Fluorescence
  • Guanosine / analogs & derivatives*
  • Inosine* / analogs & derivatives
  • Inosine* / metabolism
  • Kinetics

Substances

  • Enzyme Inhibitors
  • Guanosine
  • Inosine
  • Adenosine Deaminase
  • Adenosine