Glycoproteomic characterization of recombinant mouse α-dystroglycan.

Rebecca Harrison; Paul G. Hitchen; Maria Panico; Howard R. Morris; David Mekhaiel; Richard Pleass; Anne Dell; Jane E. Hewitt; Stuart M. Haslam

doi:10.1093/glycob/cws002

Glycoproteomic characterization of recombinant mouse α-dystroglycan.

Rebecca Harrison, Paul G. Hitchen, Maria Panico, Howard R. Morris, David Mekhaiel, Richard Pleass, Anne Dell, Jane E. Hewitt, Stuart M. Haslam

Tropical Disease Biology

Research output: Contribution to journal › Article › peer-review

52 Citations (Scopus)

Abstract

α-Dystroglycan (DG) is a key component of the dystrophin-glycoprotein complex. Aberrant glycosylation of the protein has been linked to various forms of congenital muscular dystrophy. Unusually α-DG has previously been demonstrated to be modified with both O-N-acetylgalactosamine and O-mannose initiated glycans. In the present study, Fc-tagged recombinant mouse α-DG was expressed and purified from human embryonic kidney 293T cells. α-DG glycopeptides were characterized by glycoproteomic strategies using both nano-liquid chromatography matrix-assisted laser desorption ionization and electrospray tandem mass spectrometry. A total of 14 different peptide sequences and 38 glycopeptides were identified which displayed heterogeneous O-glycosylation. These data provide new insights into the complex domain-specific O-glycosylation of α-DG.

Original language	English
Pages (from-to)	662-675
Number of pages	14
Journal	Glycobiology
Volume	22
Issue number	5
DOIs	https://doi.org/10.1093/glycob/cws002
Publication status	Published - 1 May 2012

Keywords

Dystroglycan
glycoproteomics
mass spectrometry
O-GalNAc
O-mannose

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10.1093/glycob/cws002

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title = "Glycoproteomic characterization of recombinant mouse α-dystroglycan.",

abstract = "α-Dystroglycan (DG) is a key component of the dystrophin-glycoprotein complex. Aberrant glycosylation of the protein has been linked to various forms of congenital muscular dystrophy. Unusually α-DG has previously been demonstrated to be modified with both O-N-acetylgalactosamine and O-mannose initiated glycans. In the present study, Fc-tagged recombinant mouse α-DG was expressed and purified from human embryonic kidney 293T cells. α-DG glycopeptides were characterized by glycoproteomic strategies using both nano-liquid chromatography matrix-assisted laser desorption ionization and electrospray tandem mass spectrometry. A total of 14 different peptide sequences and 38 glycopeptides were identified which displayed heterogeneous O-glycosylation. These data provide new insights into the complex domain-specific O-glycosylation of α-DG.",

keywords = "Dystroglycan, glycoproteomics, mass spectrometry, O-GalNAc, O-mannose",

author = "Rebecca Harrison and Hitchen, \{Paul G.\} and Maria Panico and Morris, \{Howard R.\} and David Mekhaiel and Richard Pleass and Anne Dell and Hewitt, \{Jane E.\} and Haslam, \{Stuart M.\}",

year = "2012",

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doi = "10.1093/glycob/cws002",

language = "English",

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T1 - Glycoproteomic characterization of recombinant mouse α-dystroglycan.

AU - Harrison, Rebecca

AU - Hitchen, Paul G.

AU - Panico, Maria

AU - Morris, Howard R.

AU - Mekhaiel, David

AU - Pleass, Richard

AU - Dell, Anne

AU - Hewitt, Jane E.

AU - Haslam, Stuart M.

PY - 2012/5/1

Y1 - 2012/5/1

N2 - α-Dystroglycan (DG) is a key component of the dystrophin-glycoprotein complex. Aberrant glycosylation of the protein has been linked to various forms of congenital muscular dystrophy. Unusually α-DG has previously been demonstrated to be modified with both O-N-acetylgalactosamine and O-mannose initiated glycans. In the present study, Fc-tagged recombinant mouse α-DG was expressed and purified from human embryonic kidney 293T cells. α-DG glycopeptides were characterized by glycoproteomic strategies using both nano-liquid chromatography matrix-assisted laser desorption ionization and electrospray tandem mass spectrometry. A total of 14 different peptide sequences and 38 glycopeptides were identified which displayed heterogeneous O-glycosylation. These data provide new insights into the complex domain-specific O-glycosylation of α-DG.

AB - α-Dystroglycan (DG) is a key component of the dystrophin-glycoprotein complex. Aberrant glycosylation of the protein has been linked to various forms of congenital muscular dystrophy. Unusually α-DG has previously been demonstrated to be modified with both O-N-acetylgalactosamine and O-mannose initiated glycans. In the present study, Fc-tagged recombinant mouse α-DG was expressed and purified from human embryonic kidney 293T cells. α-DG glycopeptides were characterized by glycoproteomic strategies using both nano-liquid chromatography matrix-assisted laser desorption ionization and electrospray tandem mass spectrometry. A total of 14 different peptide sequences and 38 glycopeptides were identified which displayed heterogeneous O-glycosylation. These data provide new insights into the complex domain-specific O-glycosylation of α-DG.

KW - Dystroglycan

KW - glycoproteomics

KW - mass spectrometry

KW - O-GalNAc

KW - O-mannose

U2 - 10.1093/glycob/cws002

DO - 10.1093/glycob/cws002

M3 - Article

SN - 0959-6658

VL - 22

SP - 662

EP - 675

JO - Glycobiology

JF - Glycobiology

IS - 5

ER -

Glycoproteomic characterization of recombinant mouse α-dystroglycan.

Abstract

Keywords

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