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

DOI: 10.1021/jacs.0c13059

Authors: Yurong Chen (Leiden University) , Zachary Armstrong (University of York) , Marta Artola (Leiden University) , Bogdan I. Florea (Leiden University) , Chi-Lin Kuo (Leiden University) , Casper De Boer (Leiden University) , Mikkel S. Rasmussen (Technical University of Denmark) , Maher Abou Hachem (echnical University of Denmark) , Gijsbert A. Van Der Marel (Leiden University) , Jeroen D. C. Codée (Leiden University) , Johannes M. F. G. Aerts (Leiden University) , Gideon J. Davies (University of York) , Herman S. Overkleeft (Leiden University)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Journal Of The American Chemical Society , VOL 346

State: Published (Approved)
Published: January 2021
Diamond Proposal Number(s): 18598

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.

Journal Keywords: Peptides and proteins; Carbohydrates; Labeling; Inhibitors; Proteomics

Diamond Keywords: Enzymes

Subject Areas: Biology and Bio-materials, Chemistry


Instruments: I03-Macromolecular Crystallography , I04-Macromolecular Crystallography

Added On: 01/02/2021 10:10

Documents:
chen-et-al-2021-activity-based-protein-profiling-of-retaining-%CE%B1-amylases-in-complex-biological-samples.pdf

Discipline Tags:

Biotechnology Biochemistry Catalysis Chemistry Structural biology Engineering & Technology Organic Chemistry Life Sciences & Biotech

Technical Tags:

Diffraction Macromolecular Crystallography (MX)