The
dystrophin glycoprotein complex, which connects the cell membrane to the basement membrane, is essential for a variety of biological events, including maintenance of muscle integrity. An O-
mannose-type GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man structure of α-
dystroglycan (α-DG), a subunit of the complex that is anchored to the cell membrane, interacts directly with
laminin in the basement membrane. Reduced glycosylation of α-DG is linked to some types of inherited
muscular dystrophy; consistent with this relationship, many disease-related mutations have been detected in genes involved in O-mannosyl
glycan synthesis. Defects in
protein O-linked
mannose β1,2-N-acetylglucosaminyltransferase 1 (
POMGnT1), a
glycosyltransferase that participates in the formation of GlcNAc-β1,2-Man
glycan, are causally related to
muscle-eye-brain disease (MEB), a congenital
muscular dystrophy, although the role of
POMGnT1 in postphosphoryl modification of GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man
glycan remains elusive. Our crystal structures of
POMGnT1 agreed with our previous results showing that the catalytic domain recognizes substrate O-mannosylated
proteins via hydrophobic interactions with little sequence specificity. Unexpectedly, we found that the stem domain recognizes the β-linked GlcNAc of O-mannosyl
glycan, an enzymatic product of
POMGnT1. This interaction may recruit
POMGnT1 to a specific site of α-DG to promote GlcNAc-β1,2-Man clustering and also may recruit other
enzymes that interact with
POMGnT1, e.g., fukutin, which is required for further modification of the GalNAc-β1,3-GlcNAc-β1,4-(phosphate-6)-Man
glycan. On the basis of our findings, we propose a mechanism for the deficiency in postphosphoryl modification of the
glycan observed in POMGnT1-KO mice and MEB patients.