Detection of Bacterial α-<span style="font-variant: small-caps">l</span>-Fucosidases with an <i>Ortho</i>-Quinone Methide-Based Probe and Mapping of the Probe-Protein Adducts
Yvette M. C. A. Luijkx,
Anniek J. Henselijn,
Gerlof P. Bosman,
Dario A. T. Cramer,
Koen C. A. P. Giesbers,
Esther M. van ‘t Veld,
Geert-Jan Boons,
Albert J. R. Heck,
Karli R. Reiding,
Karin Strijbis,
Tom Wennekes
Affiliations
Yvette M. C. A. Luijkx
Department Chemical Biology and Drug Discovery, Utrecht Institute for Pharmaceutical Sciences, Bijvoet Center for Biomolecular Research, Utrecht University, Universiteitswg 99, 3584 CG Utrecht, The Netherlands
Anniek J. Henselijn
Department Chemical Biology and Drug Discovery, Utrecht Institute for Pharmaceutical Sciences, Bijvoet Center for Biomolecular Research, Utrecht University, Universiteitswg 99, 3584 CG Utrecht, The Netherlands
Gerlof P. Bosman
Department Chemical Biology and Drug Discovery, Utrecht Institute for Pharmaceutical Sciences, Bijvoet Center for Biomolecular Research, Utrecht University, Universiteitswg 99, 3584 CG Utrecht, The Netherlands
Dario A. T. Cramer
Department Biomolecular Mass Spectrometry and Proteomics, Utrecht Institute for Pharmaceutical Sciences, Bijvoet Center for Biomolecular Research, Utrecht University, 3584 CH Utrecht, The Netherlands
Koen C. A. P. Giesbers
Department Biomolecular Health Sciences, Division Infectious Diseases and Immunology, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 1, 3584 CL Utrecht, The Netherlands
Esther M. van ‘t Veld
Department Biomolecular Health Sciences, Centre for Cell Imaging, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 1, 3584 CL Utrecht, The Netherlands
Geert-Jan Boons
Department Chemical Biology and Drug Discovery, Utrecht Institute for Pharmaceutical Sciences, Bijvoet Center for Biomolecular Research, Utrecht University, Universiteitswg 99, 3584 CG Utrecht, The Netherlands
Albert J. R. Heck
Department Biomolecular Mass Spectrometry and Proteomics, Utrecht Institute for Pharmaceutical Sciences, Bijvoet Center for Biomolecular Research, Utrecht University, 3584 CH Utrecht, The Netherlands
Karli R. Reiding
Department Biomolecular Mass Spectrometry and Proteomics, Utrecht Institute for Pharmaceutical Sciences, Bijvoet Center for Biomolecular Research, Utrecht University, 3584 CH Utrecht, The Netherlands
Karin Strijbis
Department Biomolecular Health Sciences, Division Infectious Diseases and Immunology, Faculty of Veterinary Medicine, Utrecht University, Yalelaan 1, 3584 CL Utrecht, The Netherlands
Tom Wennekes
Department Chemical Biology and Drug Discovery, Utrecht Institute for Pharmaceutical Sciences, Bijvoet Center for Biomolecular Research, Utrecht University, Universiteitswg 99, 3584 CG Utrecht, The Netherlands
Fucosidases are associated with several pathological conditions and play an important role in the health of the human gut. For example, fucosidases have been shown to be indicators and/or involved in hepatocellular carcinoma, breast cancer, and helicobacter pylori infections. A prerequisite for the detection and profiling of fucosidases is the formation of a specific covalent linkage between the enzyme of interest and the activity-based probe (ABP). The most commonly used fucosidase ABPs are limited to only one of the classes of fucosidases, the retaining fucosidases. New approaches are needed that allow for the detection of the second class of fucosidases, the inverting type. Here, we report an ortho-quinone methide-based probe with an azide mini-tag that selectively labels both retaining and inverting bacterial α-l-fucosidases. Mass spectrometry-based intact protein and sequence analysis of a probe-labeled bacterial fucosidase revealed almost exclusive single labeling at two specific tryptophan residues outside of the active site. Furthermore, the probe could detect and image extracellular fucosidase activity on the surface of live bacteria.
