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Instruments / Technical Area	Publication Details	Published	Actions
I03-Macromolecular Crystallography I04-Macromolecular Crystallography	Activity-based protein profiling of retaining α-amylases in complex biological samples Yurong Chen , Zachary Armstrong , Marta Artola , Bogdan I. Florea , Chi-Lin Kuo , Casper De Boer , Mikkel S. Rasmussen , Maher Abou Hachem , Gijsbert A. Van Der Marel , Jeroen D. C. Codée , Johannes M. F. G. Aerts , Gideon J. Davies , Herman S. Overkleeft Journal Of The American Chemical Society 346 DOI: 10.1021/jacs.0c13059 Diamond Proposal Number(s): [18598] Open Access Show Abstract ▼ Abstract: Amylases are key enzymes in the processing of starch in many kingdoms of life. They are important catalysts in industrial biotechnology where they are applied in, among others, food processing and the production of detergents. In man amylases are the first enzymes in the digestion of starch to glucose and arguably also the preferred target in therapeutic strategies aimed at the treatment of type 2 diabetes patients through down-tuning glucose assimilation. Efficient and sensitive assays that report selectively on retaining amylase activities irrespective of the nature and complexity of the biomaterial studied are of great value both in finding new and effective human amylase inhibitors and in the discovery of new microbial amylases with potentially advantageous features for biotechnological application. Activity-based protein profiling (ABPP) of retaining glycosidases is inherently suited for the development of such an assay format. We here report on the design and synthesis of 1,6-epi-cyclophellitol-based pseudodisaccharides equipped with a suite of reporter entities and their use in ABPP of retaining amylases from human saliva, murine tissue as well as secretomes from fungi grown on starch. The activity and efficiency of the inhibitors and probes are substantiated by extensive biochemical analysis, and the selectivity for amylases over related retaining endoglycosidases is validated by structural studies.	Jan 2021

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I03-Macromolecular Crystallography
I04-Macromolecular Crystallography

Activity-based protein profiling of retaining α-amylases in complex biological samples