Glycoproteomic characterization of recombinant mouse α-dystroglycan

alpha-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 alpha-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 alpha-DG was expressed and purified from human embryonic kidney 293T cells. alpha-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 glycosylation. These data provide new insights into the complex domain-specific and 38 glycopeptides were identified which displayed heterogeneous O-O-glycosylation of alpha-DG.