TY - JOUR
T1 - Ligand-Directed Chemistry on Glycoside Hydrolases - A Proof of Concept Study
AU - Prasch, Herwig Anton
AU - Wolfsgruber, Andreas
AU - Thonhofer, Martin Simon
AU - Culum, Andre Stephan
AU - Mandl, Christoph
AU - Zündel, Melanie
AU - Weber, Patrick
AU - Nasseri, Seyed A.
AU - Gonzales-Santana, Andrés
AU - Tegl, Gregor
AU - Nidetzky, Bernd
AU - Gruber, Karl
AU - Stütz, Arnold
AU - Withers, Stephen G
AU - Wrodnigg, Tanja Maria
PY - 2023/12/1
Y1 - 2023/12/1
N2 - 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.
AB - 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.
KW - Ligand-directed chemistry
KW - iminoalditol probes
KW - glycoside hydrolases
KW - beta-glucosidases
KW - covalent enzyme labelling
KW - covalent enzyme labelling
KW - glycoside hydrolases
KW - iminoalditol probes
KW - ligand-directed chemistry
KW - β-glucosidases
UR - http://www.scopus.com/inward/record.url?scp=85173781498&partnerID=8YFLogxK
U2 - 10.1002/cbic.202300480
DO - 10.1002/cbic.202300480
M3 - Article
SN - 1439-4227
VL - 24
JO - ChemBioChem
JF - ChemBioChem
IS - 23
M1 - e202300480
ER -