title: LARGE expression augments the glycosylation of glycoproteins in addition to α-dystroglycan conferring laminin binding.
abstract: Mutations in genes encoding glycosyltransferases (and presumed glycosyltransferases) that affect glycosylation and extracellular matrix binding activity of α-dystroglycan (α-DG) cause congenital muscular dystrophies (CMDs) with central nervous system manifestations. Among the identified genes, LARGE is of particular interest because its overexpression rescues glycosylation defects of α-DG in mutations of not only LARGE but also other CMD-causing genes and restores laminin binding activity of α-DG. It is not known whether LARGE protein glycosylates other proteins in addition to α-DG. In this study, we overexpressed LARGE in DG-deficient cells and analyzed glycosylated proteins by Western blot analysis. Surprisingly, overexpression of LARGE in α-DG-deficient cells led to glycosylation dependent IIH6C4 and VIA4-1 immunoreactivity, despite the prevailing view that these antibodies only recognize glycosylated α-DG. Furthermore, the hyperglycosylated proteins in LARGE-overexpressing cells demonstrated the functional capacity to bind the extracellular matrix molecule laminin and promote laminin assembly at the cell surface, an effect that was blocked by IIH6C4 antibodies. These results indicate that overexpression of LARGE catalyzes the glycosylation of at least one other glycoprotein in addition to α-DG, and that this glycosylation(s) promotes laminin binding activity.
authors.affiliation: Department of Neuroscience and Physiology, Upstate Medical University, Syracuse, New York, United States of America.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Zhang
authors.forename: Peng
authors.initials: P
authors.lastname: Zhang
authors.forename: Huaiyu
authors.initials: H
authors.lastname: Hu
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D002478:Cells, Cultured; D017931:DNA Primers; D049030:Dystroglycans / Q000235:genetics / Q000378:metabolism*; D055786:Gene Knockout Techniques; D006023:Glycoproteins / Q000378:metabolism*; D006031:Glycosylation; D007797:Laminin / Q000378:metabolism*; D051379:Mice; D008822:Mice, Transgenic; D017351:N-Acetylglucosaminyltransferases / Q000235:genetics*; D009474:Neurons / Q000378:metabolism; D011485:Protein Binding; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013234:Stem Cells / Q000378:metabolism
chemical_list: D017931:DNA Primers; D006023:Glycoproteins; D007797:Laminin; D049030:Dystroglycans; C502747:Large1 protein, mouse; D017351:N-Acetylglucosaminyltransferases