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Recent advances and concepts in substrate specificity determination of proteases using tailored libraries of fluorogenic substrates with unnatural amino acids

  • Wioletta Rut , Paulina Kasperkiewicz , Anna Byzia , Marcin Poreba , Katarzyna Groborz and Marcin Drag EMAIL logo
Published/Copyright: February 21, 2015

Abstract

Substrate specificity of proteases can be determined using several methods among which the most frequently used are positional scanning library, proteomics and phage display. Classic approaches can deliver information about preferences for natural amino acids in binding pockets of virtually all proteases. However, recent studies demonstrate the ability to obtain much more information by application of unnatural amino acids to positional scanning library approaches. This knowledge can be used for the design of more active and specific substrates, inhibitors and activity based probes. In this minireview we describe recent strategies and concepts for the design and application of fluorogenic substrates library tailored for exopeptidases and endopeptidases.


Corresponding author: Marcin Drag, Division of Bioorganic Chemistry, Faculty of Chemistry, Wroclaw University of Technology, Wyb. Wyspianskiego 27, Wroclaw 50-370, Poland, e-mail:
aThese authors contributed equally to this article.

Acknowledgments

This work was supported by the National Science Centre grant 2011/03/B/ST5/01048 in Poland. The work was also supported by a statutory activity subsidy from the Polish Ministry of Science and Higher Education for the Faculty of Chemistry at Wroclaw University of Technology. We would like to thank Guy S. Salvesen for critical reading of the manuscript.

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Received: 2014-12-16
Accepted: 2015-1-27
Published Online: 2015-2-21
Published in Print: 2015-4-1

©2015 by De Gruyter

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