5I23

Crystal Structure of Agd31B, alpha-transglucosylase in Glycoside Hydrolase Family 31, in complex with Cyclophellitol Aziridine probe CF022

Summary for 5I23

• Entry DOI: 10.2210/pdb5i23/pdb
• Descriptor: Oligosaccharide 4-alpha-D-glucosyltransferase, TETRAETHYLENE GLYCOL, SULFATE ION, ... (6 entities in total)
• Functional Keywords: alpha glycosidase, cyclophellitol aziridine, inhibitor, probe, transferase
• Biological source: Cellvibrio japonicus (strain Ueda107)
• Total number of polymer chains: 1
• Total formula weight: 95834.87

Authors

Wu, L.,Davies, G.J. (deposition date: 2016-02-08, release date: 2016-05-04, Last modification date: 2024-01-10)

Primary citation

Jiang, J.,Kuo, C.L.,Wu, L.,Franke, C.,Kallemeijn, W.W.,Florea, B.I.,van Meel, E.,van der Marel, G.A.,Codee, J.D.,Boot, R.G.,Davies, G.J.,Overkleeft, H.S.,Aerts, J.M.
Detection of Active Mammalian GH31 alpha-Glucosidases in Health and Disease Using In-Class, Broad-Spectrum Activity-Based Probes.
Acs Cent.Sci., 2:351-358, 2016

PubMed Abstract: The development of small molecule activity-based probes (ABPs) is an evolving and powerful area of chemistry. There is a major need for synthetically accessible and specific ABPs to advance our understanding of enzymes in health and disease. α-Glucosidases are involved in diverse physiological processes including carbohydrate assimilation in the gastrointestinal tract, glycoprotein processing in the endoplasmic reticulum (ER), and intralysosomal glycogen catabolism. Inherited deficiency of the lysosomal acid α-glucosidase (GAA) causes the lysosomal glycogen storage disorder, Pompe disease. Here, we design a synthetic route for fluorescent and biotin-modified ABPs for in vitro and in situ monitoring of α-glucosidases. We show, through mass spectrometry, gel electrophoresis, and X-ray crystallography, that α-glucopyranose configured cyclophellitol aziridines label distinct retaining α-glucosidases including GAA and ER α-glucosidase II, and that this labeling can be tuned by pH. We illustrate a direct diagnostic application in Pompe disease patient cells, and discuss how the probes may be further exploited for diverse applications.

PubMed: 27280170
DOI: 10.1021/acscentsci.6b00057

Experimental method

X-RAY DIFFRACTION (1.95 Å)