Department of Cell Biology and Infection
Department of Structural Biology and Chemistry
Tuesday 22 october 2019 at 2.00 pm
AUDITORIUM CENTRE F. JACOB room- CFJ RdC 17
Leiden Institute of Chemistry, Leiden University
Activity-based glycosidase profiling in biomedicine and biotechnology
Activity-based protein profiling (ABPP) is a rapidly emerging field in chemical biology research. Enzymes that employ a mechanism in processing their substrate that involves formation of a covalent enzyme-intermediate adduct can be blocked by mechanism-based suicide inhibitors: compounds that react within the enzyme active site to form a covalent and irreversible adduct. Introduction of a reporter moiety (‘TAG’ in the below picture) yields an activity-based probe (ABP) through which enzyme activities can be discovered (comparative ABPP) and the efficacy enzyme inhibitors in complex biological systems analyzed (competitive ABPP).
Our work on ABPP development focuses on retaining glycosidases: hydrolytic enzymes able to cleave interglycosidic linkages and that do so through the formation of covalent enzymesubstrate intermediates. Configurational and functional analogues of the natural product and mechanism-based retaining beta-glucosidase inhibitor, cyclophellitol, prove to be highly versatile tools to study retaining glycosidases of various nature and origin in relation to human health and disease, but also in the field of biotechnology. In this lecture the current state in the design, synthesis and application of synthetic cyclophellitol derivatives in studying retaining glycosidases will be presented. Discussed subjects will include 1) diagnosis of human lysosomal exoglycosidases in relation to lysosomal storage disorders; 2) glycosylation of cyclophellitol derivatives top arrive at retaining endoglycosidase ABPs and 3) application of glycosidase ABPs in the functional profiling of fungal secretomes for the discovery of glycosidases for biotechnology application.
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3) L. Wu, J. Jiang, Y. Jin, W. W. Kallemeijn, C.-L. Kuo, M. Artola, W. Dai, C. van Elk, M. van Eijk, G. A. van der Marel, J. D. C. Codée, B. I. Florea, J. M. F. G. Aerts, H. S. Overkleeft and G. J. Davies, Activity-based probes forfunctional interrogation of retaining beta-glucuronidases, Nat. Chem. Biol. 2017, 13, 867-873.
4) S. P. Schröder, C. de Boer, N. G. S. McGregor, R. J. Rowland, O. Moroz, E. Blagova, J. Reijngoud, M. Arentshorst, D. Osborn, M. D. Morant, E. Abbate, M. A. Stringer, K. B. R. M. Krogh, L. Raich, C. Rovira, J.-G. Berrin, G. P. van Wezel, A. F. J. Ram, B. I. Florea, G. A. van der Marel, J. D. C. Codée, K. S. Wilson, L. Wu, G. J. Davies and H. S. Overkleeft, Dynamic and functional profiling of xylan-degrading enzymes in Aspergillus secretomes using activity-based probes, ACS Cent. Sci 2019, 5, 1067-1078.
Unité de Pathogénèse des infections vasculaires
Unité Chimie Biologique Epigénétique