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

TY - JOUR

T1 - Detection of Active Mammalian GH31 α-Glucosidases in Health and Disease Using In-Class, Broad-Spectrum Activity-Based Probes

AU - Jiang, Jianbing

AU - Kuo, Chi-Lin

AU - Wu, Liang

AU - Franke, Christian

AU - Kallemeijn, Wouter W

AU - Florea, Bogdan I

AU - van Meel, Eline

AU - van der Marel, Gijsbert A

AU - Codée, Jeroen D C

AU - Boot, Rolf G

AU - Davies, Gideon J

AU - Overkleeft, Herman S

AU - Aerts, Johannes M F G

N1 - © 2016, American Chemical Society. Uploaded with permission of the publisher/copyright holder. Further copying may not be permitted; contact the publisher for details.

PY - 2016/5/25

Y1 - 2016/5/25

N2 - 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.

AB - 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.

KW - Journal Article

UR - http://www.scopus.com/inward/record.url?scp=85020875956&partnerID=8YFLogxK

U2 - 10.1021/acscentsci.6b00057

DO - 10.1021/acscentsci.6b00057

M3 - Article

C2 - 27280170

SN - 2374-7943

VL - 2

SP - 351

EP - 358

JO - ACS Central Science

JF - ACS Central Science

IS - 5

ER -