A mass spectrometry plate reader: Monitoring enzyme activity and inhibition with a desorption/ionization on silicon (DIOS) platform

Zhouxin Shen, Eden P. Go, Alejandra Gamez, Junefredo V. Apon, Valery Fokin, Mike Greig, Manuel Ventura, John E. Crowell, Ola Blixt, James C. Paulson, Raymond C. Stevens, M. G. Finn, Gary Siuzdak

Research output: Contribution to journalArticle

65 Citations (Scopus)

Abstract

A surface-based laser desorption/ionization mass spectrometry assay that makes use of Desorption/Ionization on Silicon Mass Spectrometry (DIOS-MS) has been developed to monitor enzyme activity and enzyme inhibition. DIOS-MS has been used to characterize inhibitors from a library and then to monitor their activity against selected enzyme targets, including proteases, glycotransferase, and acetylcholinesterase. An automated DIOS-MS system was also used as a high-throughput screen for the activity of novel enzymes and enzyme inhibitors. On two different commercially available instruments, a sampling rate of up to 38 inhibitors per minute was accomplished, with thousands of inhibitors being monitored. The ease of applying mass spectrometry toward developing enzyme assays and the speed of surface-based assays such as DIOS for monitoring inhibitor effectiveness and enzyme activity makes it attractive for a broad range of screening applications.

Original languageEnglish
Pages (from-to)921-927
Number of pages7
JournalChemBioChem
Volume5
Issue number7
DOIs
Publication statusPublished - 5 Jul 2004
Externally publishedYes

Keywords

  • DIOS
  • Enzymes
  • High-throughput screening
  • Inhibitors
  • Plate reader

ASJC Scopus subject areas

  • Drug Discovery
  • Organic Chemistry
  • Biochemistry, Genetics and Molecular Biology(all)

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    Shen, Z., Go, E. P., Gamez, A., Apon, J. V., Fokin, V., Greig, M., Ventura, M., Crowell, J. E., Blixt, O., Paulson, J. C., Stevens, R. C., Finn, M. G., & Siuzdak, G. (2004). A mass spectrometry plate reader: Monitoring enzyme activity and inhibition with a desorption/ionization on silicon (DIOS) platform. ChemBioChem, 5(7), 921-927. https://doi.org/10.1002/cbic.200400008