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Glycomic Analyses of Mouse Models of Congenital Muscular Dystrophy
Journal article   Open access   Peer reviewed

Glycomic Analyses of Mouse Models of Congenital Muscular Dystrophy

Stephanie H Stalnaker, Kazuhiro Aoki, Jae-Min Lim, Mindy Porterfield, Mian Liu, Jakob S Satz, Sean Buskirk, Yufang Xiong, Peng Zhang, Kevin P Campbell, …
The Journal of biological chemistry, Vol.286(24), pp.21180-21190
06/17/2011
DOI: 10.1074/jbc.M110.203281
PMCID: PMC3122180
PMID: 21460210
url
https://doi.org/10.1074/jbc.M110.203281View
Published (Version of record) Open Access

Abstract

Dystroglycanopathies are a subset of congenital muscular dystrophies wherein α-dystroglycan (α-DG) is hypoglycosylated. α-DG is an extensively O -glycosylated extracellular matrix-binding protein and a key component of the dystrophin-glycoprotein complex. Previous studies have shown α-DG to be post-translationally modified by both O -GalNAc- and O -mannose-initiated glycan structures. Mutations in defined or putative glycosyltransferase genes involved in O -mannosylation are associated with a loss of ligand-binding activity of α-DG and are causal for various forms of congenital muscular dystrophy. In this study, we sought to perform glycomic analysis on brain O -linked glycan structures released from proteins of three different knock-out mouse models associated with O -mannosylation (POMGnT1, LARGE (Myd), and DAG1 −/− ). Using mass spectrometry approaches, we were able to identify nine O -mannose-initiated and 25 O -GalNAc-initiated glycan structures in wild-type littermate control mouse brains. Through our analysis, we were able to confirm that POMGnT1 is essential for the extension of all observed O -mannose glycan structures with β1,2-linked GlcNAc. Loss of LARGE expression in the Myd mouse had no observable effect on the O -mannose-initiated glycan structures characterized here. Interestingly, we also determined that similar amounts of O -mannose-initiated glycan structures are present on brain proteins from α-DG-lacking mice (DAG1) compared with wild-type mice, indicating that there must be additional proteins that are O -mannosylated in the mammalian brain. Our findings illustrate that classical β1,2-elongation and β1,6-GlcNAc branching of O -mannose glycan structures are dependent upon the POMGnT1 enzyme and that O -mannosylation is not limited solely to α-DG in the brain.
Carbohydrate-binding Protein POMGnT1 Glycomics LARGE Glycoprotein Carbohydrate Function Mass Spectrometry (MS) Glycobiology and Extracellular Matrices alpha-Dystroglycan Muscular Dystrophy Carbohydrate Structure

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