Skip to main content

Novel method for detection of glycogen in cells.

Citation
Skurat, A., et al. “Novel Method For Detection Of Glycogen In Cells.”. Glycobiology, pp. 416-424.
Center Indiana University
Author Alexander Skurat V, Dyann M Segvich, Anna A DePaoli-Roach, Peter J Roach
Keywords CBM20, Lafora disease, Pompe disease, glycogen, immunofluorescence
Abstract

y: Glycogen, a branched polymer of glucose, functions as an energy reserve in many living organisms. Abnormalities in glycogen metabolism, usually excessive accumulation, can be caused genetically, most often through mutation of the enzymes directly involved in synthesis and degradation of the polymer leading to a variety of glycogen storage diseases (GSDs). Microscopic visualization of glycogen deposits in cells and tissues is important for the study of normal glycogen metabolism as well as diagnosis of GSDs. Here, we describe a method for the detection of glycogen using a renewable, recombinant protein which contains the carbohydrate-binding module (CBM) from starch-binding domain containing protein 1 (Stbd1). We generated a fusion protein containing g lutathione S-transferase, a cM c eptitope and the tbd1 BM (GYSC) for use as a glycogen-binding probe, which can be detected with secondary antibodies against glutathione S-transferase or cMyc. By enzyme-linked immunosorbent assay, we demonstrate that GYSC binds glycogen and two other polymers of glucose, amylopectin and amylose. Immunofluorescence staining of cultured cells indicate a GYSC-specific signal that is co-localized with signals obtained with anti-glycogen or anti-glycogen synthase antibodies. GYSC-positive staining inside of lysosomes is observed in individual muscle fibers isolated from mice deficient in lysosomal enzyme acid alpha-glucosidase, a well-characterized model of GSD II (Pompe disease). Co-localized GYSC and glycogen signals are also found in muscle fibers isolated from mice deficient in malin, a model for Lafora disease. These data indicate that GYSC is a novel probe that can be used to study glycogen metabolism under normal and pathological conditions.

Year of Publication
2017
Journal
Glycobiology
Volume
27
Issue
5
Number of Pages
416-424
Date Published
12/2017
ISSN Number
1460-2423
DOI
10.1093/glycob/cwx005
Alternate Journal
Glycobiology
PMID
28077463
PMCID
PMC5444244
Download citation