Detection of Active Mammalian GH31 α-Glucosidases in Health and Disease Using In-Class, Broad-Spectrum Activity-Based Probes

DOI: 10.1021/acscentsci.6b00057

Authors: Jianbing Jiang (Department of Bio-organic Synthesis, Leiden Institute of Chemistry, Leiden University) , Chi-lin Kuo (Department of Medical Biochemistry, Leiden Institute of Chemistry, Leiden University) , Liang Wu (University of York) , Christian Franke (Department of Bio-organic Synthesis, Leiden Institute of Chemistry, Leiden University) , Wouter W. Kallemeijn (Department of Medical Biochemistry, Leiden Institute of Chemistry, Leiden University) , Bogdan I. Florea (Department of Bio-organic Synthesis, Leiden Institute of Chemistry, Leiden University) , Eline Van Meel (Department of Medical Biochemistry, Leiden Institute of Chemistry, Leiden University) , Gijsbert A. Van Der Marel (Department of Bio-organic Synthesis, Leiden Institute of Chemistry, Leiden University) , Jeroen D. C. Codée (Department of Bio-organic Synthesis, Leiden Institute of Chemistry, Leiden University) , Rolf G. Boot (Department of Medical Biochemistry, Leiden Institute of Chemistry, Leiden University) , Gideon J. Davies (University of York) , Herman S. Overkleeft (Department of Bio-organic Synthesis, Leiden Institute of Chemistry, Leiden University) , Johannes M. F. G. Aerts (Department of Medical Biochemistry, Leiden Institute of Chemistry, Leiden University)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Acs Central Science , VOL 2 , PAGES 351 - 358

State: Published (Approved)
Published: May 2016
Diamond Proposal Number(s): 9948

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.

Subject Areas: Biology and Bio-materials, Chemistry


Instruments: I04-Macromolecular Crystallography

Documents:
acscentsci.pdf