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

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Author(s) Bovet, Cédric, Zenobi, Renato
Publication Type Journal Items, Publication Status: Published
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Title Determination of active enzyme concentration using activity-based probes and direct mass spectrometric readout
Author(s) Bovet, Cédric
Zenobi, Renato
Journal or Series Title Analytical biochemistry
Volume Number 373
Issue Number 2
Start Page 380
End Page 382
ISSN 0003-2697
Publisher Elsevier
Publication Place San Diego
Publication Date 2008
Abstract 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.
DOI 10.1016/j.ab.2007.11.003
Additional Notes Received 25 July 2007, Available online 6 November 2007
Document Type Article
Publication Status Published
Language English
Assigned Organisational Unit(s) 03430
Organisational Unit(s)
NEBIS System Number 000032007
Source Database ID PP-40963
WOS-000252514100024
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@article{Bvt2008,
  author = "Bovet, C{\'{e}}dric and Zenobi, Renato",
  title = "{D}etermination of active enzyme concentration using activity-based probes and direct mass spectrometric readout",
  journal = "Analytical biochemistry",
  year = 2008,
  volume = "373",
  number = "2",
  pages = "380--382",
}


E-Citations record created: Thu, 01 Apr 2010, 22:44:02 CET