Ligand-Directed Chemistry on Glycoside Hydrolases – A Proof of Concept Study |
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Authors: | Herwig Prasch Dr Andreas Wolfsgruber Dr Martin Thonhofer André Culum Christoph Mandl Dr Patrick Weber Melanie Zündel Dr Seyed A Nasseri Dr Andres Gonzalez Santana Dr Gregor Tegl Prof?Dr Bernd Nidetzky Prof?Dr Karl Gruber Prof?Dr Arnold E Stütz Prof?Dr Stephen G Withers Prof?Dr Tanja M Wrodnigg |
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Affiliation: | 1. Graz University of Technology, Institute of Chemistry and Technology of Biobased Systems, Stremayrgasse 9, 8010 Graz, Austria;2. University of British Columbia, Department of Chemistry, 2036 Main Mall, Vancouver, BC, V6T 1Z1 Canada;3. Graz University of Technology, Institute of Biotechnology and Biochemical Engineering, Petersgasse 10-12/I, 8010 Graz, Austria;4. University of Graz, Institute of Molecular Bioscience, Humboldtstraße 50/III, 8010 Graz, Austria |
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Abstract: | Selective covalent labelling of enzymes using small molecule probes has advanced the scopes of protein profiling. The covalent bond formation to a specific target is the key step of activity-based protein profiling (ABPP), a method which has become an indispensable tool for measuring enzyme activity in complex matrices. With respect to carbohydrate processing enzymes, strategies for ABPP so far involve labelling the active site of the enzyme, which results in permanent loss of activity. Here, we report in a proof of concept study the use of ligand-directed chemistry (LDC) for labelling glycoside hydrolases near – but not in – the active site. During the labelling process, the competitive inhibitor is cleaved from the probe, departs the active site and the enzyme maintains its catalytic activity. To this end, we designed a building block synthetic concept for small molecule probes containing iminosugar-based reversible inhibitors for labelling of two model β-glucosidases. The results indicate that the LDC approach can be adaptable for covalent proximity labelling of glycoside hydrolases. |
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Keywords: | β-glucosidases covalent enzyme labelling glycoside hydrolases iminoalditol probes ligand-directed chemistry |
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