Mass-Spectrometry-Linked Screening of Protein Fractions for Enzymatic Activities—A Tool for Functional Genomics
✍ Scribed by Joachim Jankowski; Nina Stephan; Martin Knobloch; Sven Fischer; Dominik Schmaltz; Walter Zidek; Hartmut Schlüter
- Publisher
- Elsevier Science
- Year
- 2001
- Tongue
- English
- Weight
- 94 KB
- Volume
- 290
- Category
- Article
- ISSN
- 0003-2697
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✦ Synopsis
A simple and rapid strategy is described to screen protein fractions for defined enzymatic activity. A protein fraction from a porcine kidney extract was immobilized by covalent coupling to activated affinity beads. The immobilized proteins were incubated with probes specific for different enzyme activities. The reaction products were analyzed by matrix-assisted laser desorption/ionization (MALDI)-mass spectrometry. The MALDI spectra indicate the presence of 5-nucleotidase, phosphatase, kinase, glutathione reductase, and renin activities in the kidney protein extract. Furthermore, the method can be used to screen for inhibitors of enzymatic reactions. The method is adaptable to high-throughput sample handling and automated mass spectrometric analysis and therefore suited for functional genomics.