Dystroglycan is a highly glycosylated extracellular matrix receptor with essential functions in skeletal muscle and the nervous system. Reduced matrix binding by α-dystroglycan (α-DG) due to perturbed glycosylation is a pathological feature of several forms of muscular dystrophy. Like-acetylglucosaminyltransferase (LARGE) synthesizes the matrix-binding heteropolysaccharide [-glucuronic acid-β1,3-xylose-α1,3-]n. Using a dual exoglycosidase digestion, we confirm that this polysaccharide is present on native α-DG from skeletal muscle. The atomic details of matrix binding were revealed by a high-resolution crystal structure of laminin-G-like (LG) domains 4 and 5 (LG4 and LG5) of laminin-α2 bound to a LARGE-synthesized oligosaccharide. A single glucuronic acid-β1,3-xylose disaccharide repeat straddles a Ca2+ ion in the LG4 domain, with oxygen atoms from both sugars replacing Ca2+-bound water molecules. The chelating binding mode accounts for the high affinity of this protein–carbohydrate interaction. These results reveal a previously uncharacterized mechanism of carbohydrate recognition and provide a structural framework for elucidating the mechanisms underlying muscular dystrophy.
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We thank S.G. Withers (University of British Columbia) for a gift of T. maritima β-glucuronidase and C.M. Blaumueller for critical reading of the manuscript. The IIH6 antibody was obtained from the Developmental Studies Hybridoma Bank, University of Iowa. We acknowledge Diamond Light Source for time on beamlines I02 and I04-1 under proposal MX9424. This work was funded by a Wellcome Trust Senior Investigator Award to E.H. (101748/Z/13/Z) and a Paul D. Wellstone Muscular Dystrophy Cooperative Research Center grant to K.P.C. (1U54NS053672).
The authors declare no competing financial interests.
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Briggs, D., Yoshida-Moriguchi, T., Zheng, T. et al. Structural basis of laminin binding to the LARGE glycans on dystroglycan. Nat Chem Biol 12, 810–814 (2016). https://doi.org/10.1038/nchembio.2146
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