Automated colorimetric screen for apyrase inhibitors

J. B. Windsor, C. Thomas, L. Hurley, S. J. Roux, Alan M. Lloyd

Research output: Contribution to journalArticle

16 Scopus citations

Abstract

Apyrases are enzymes that efficiently hydrolyze ATP and ADP and may operate both inside and outside the cell. Although apyrases are important to a variety of cellular mechanisms and uses in industry, there are no available apyrase-specific inhibitors. Colorimetric assays based on the Fiske-Subbarow method for measuring inorganic phosphate are able to detect the release of inorganic phosphate, from ATP and other nucleotides. We found that this type of assay could be automated and used to screen, for apyrase-inhibiting compounds by assaying for a reduction in released phosphate in the presence of potential inhibitors. The automation of this assay allowed for the successful screening of a commercially available compound library. Several low molecular weight compounds were identified that, when used at micromolar concentrations, effectively inhibited apyrase activity.

Original languageEnglish (US)
Pages (from-to)1024-1030
Number of pages7
JournalBioTechniques
Volume33
Issue number5
StatePublished - Nov 1 2002

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ASJC Scopus subject areas

  • Biotechnology
  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Windsor, J. B., Thomas, C., Hurley, L., Roux, S. J., & Lloyd, A. M. (2002). Automated colorimetric screen for apyrase inhibitors. BioTechniques, 33(5), 1024-1030.