LARGE2-dependent glycosylation confers laminin-binding ability on proteoglycans

Kei-Ichiro Inamori; Aaron M Beedle; Daniel Beltrán-Valero de Bernabé; Michael E Wright; Kevin P Campbell

doi:10.1093/glycob/cww075

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LARGE2-dependent glycosylation confers laminin-binding ability on proteoglycans

Journal article

Open access

Peer reviewed

LARGE2-dependent glycosylation confers laminin-binding ability on proteoglycans

Kei-Ichiro Inamori, Aaron M Beedle, Daniel Beltrán-Valero de Bernabé, Michael E Wright and Kevin P Campbell

Glycobiology, Vol.26(12), pp.1284-1296

12/2016

DOI: 10.1093/glycob/cww075

PMCID: PMC5137251

PMID: 27496765

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Abstract

Both LARGE1 (formerly LARGE) and its paralog LARGE2 are bifunctional glycosyltransferases with xylosy- and glucuronyltransferase activities, and are capable of synthesizing polymers composed of a repeating disaccharide [-3Xylα1,3GlcAβ1-]. Post-translational modification of the O-mannosyl glycan of α-dystroglycan (α-DG) with the polysaccharide is essential for it to act as a receptor for ligands in the extracellular matrix (ECM), and both LARGE paralogs contribute to the modification in vivo. LARGE1 and LARGE2 have different tissue distribution profiles and enzymatic properties; however, the functional difference of the homologs remains to be determined, and α-DG is the only known substrate for the modification by LARGE1 or LARGE2. Here we show that LARGE2 can modify proteoglycans (PGs) with the laminin-binding glycan. We found that overexpression of LARGE2, but not LARGE1, mediates the functional modification on the surface of DG , Pomt1 and Fktn embryonic stem cells. We identified a heparan sulfate-PG glypican-4 as a substrate for the LARGE2-dependent modification by affinity purification and subsequent mass spectrometric analysis. Furthermore, we showed that LARGE2 could modify several additional PGs with the laminin-binding glycan, most likely within the glycosaminoglycan (GAG)-protein linkage region. Our results indicate that LARGE2 can modify PGs with the GAG-like polysaccharide composed of xylose and glucuronic acid to confer laminin binding. Thus, LARGE2 may play a differential role in stabilizing the basement membrane and modifying its functions by augmenting the interactions between laminin globular domain-containing ECM proteins and PGs.

Embryonic Stem Cells - metabolism

Glycosyltransferases - metabolism

Animals

Glycosyltransferases - chemistry

Proteoglycans - metabolism

Laminin - chemistry

Glycosylation

Mice

Laminin - metabolism

Binding Sites

Proteoglycans - chemistry

Mice, Knockout

Details

Title: Subtitle: LARGE2-dependent glycosylation confers laminin-binding ability on proteoglycans
Creators: Kei-Ichiro Inamori - University of Iowa, Molecular Physiology and Biophysics
Aaron M Beedle - Department of Pharmaceutical and Biomedical Sciences, University of Georgia, Athens, GA 30602
Daniel Beltrán-Valero de Bernabé - Department of Neurology, Howard Hughes Medical Institute, University of Iowa Roy J. and Lucille A. Carver College of Medicine, Iowa City, IA 52242-1101, USA
Michael E Wright - University of Iowa, Molecular Physiology and Biophysics
Kevin P Campbell - University of Iowa, Molecular Physiology and Biophysics
Resource Type: Journal article
Publication Details: Glycobiology, Vol.26(12), pp.1284-1296
Event: 0
DOI: 10.1093/glycob/cww075
PMID: 27496765
PMCID: PMC5137251
NLM abbreviation: Glycobiology
ISSN: 1460-2423
eISSN: 1460-2423
Publisher: England
Grant note: U54 NS053672 / NINDS NIH HHS
Language: English
Date published: 12/2016
Academic Unit: Molecular Physiology and Biophysics; Iowa Neuroscience Institute
Record Identifier: 9983757057702771

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