Back
Determination of active enzyme concentration using activity-based probes and direct mass spectrometric readout.
Journal article

Determination of active enzyme concentration using activity-based probes and direct mass spectrometric readout.

  • Bovet C Department of Chemistry and Applied Biosciences, ETH Zurich, 8093 Zurich, Switzerland.
  • Zenobi R
  • 2007-12-11
Published in:
  • Analytical biochemistry. - 2008
Fungal Proteins
Lipase
Nanotechnology
Paraoxon
Serine Proteinase Inhibitors
Spectrometry, Mass, Electrospray Ionization
Spectrophotometry, Ultraviolet
Subtilisins
Tosyl Compounds
English Activity-based probes (ABPs) are specific covalent inhibitors developed for different classes of enzymes. We have titrated a serine protease and a lipase with their specific ABPs and measured the extent of inhibition using nanoelectrospray mass spectrometry (nanoESI-MS). Because ABPs only interact with the active enzyme form, the approach allows to accurately measure the active enzyme concentration in solution. This is even possible in the presence of contaminants. The concentrations of the two enzymes were also investigated by UV spectroscopy, which appears to give higher concentrations than those measured with the active site titration method.
Language
  • English
Open access status
closed
Identifiers
Persistent URL
https://sonar.ch/global/documents/123644
Statistics
Document views: 31 File downloads: