Structural basis of laminin binding to the LARGE glycans on dystroglycan


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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Figure 1: Enzymatic digestion of oligosaccharide G5.
Figure 2: Enzymatic digestion of skeletal-muscle α-DG.
Figure 3: Crystal structure of laminin-α2 LG4–5 bound to oligosaccharide G6/7.
Figure 4: NMR analysis of binding of oligosaccharide G5 to laminin-α2 LG4–5.
Figure 5: Structural comparison of α-DG-binding LG domains.

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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).

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D.C.B. co-designed the project, carried out the crystallographic experiments, analyzed the data and co-wrote the manuscript. T.Y.-M. produced xylosidase and glucuronidase, and performed enzyme digestions and binding assays. T.Z. generated the reagent X2, produced xylosidase and glucuronidase, and performed the G5 digestion experiment. D.V. generated reagents G3, G5 and G6/7. M.A. performed enzyme digestions and western blotting. A.S. and M.M. provided well-characterized xylosidase. L.Y. carried out the NMR experiments and analyzed the data. E.H. and K.P.C. co-designed the project, co-wrote the manuscript and supervised the research. All authors discussed the results and commented on the manuscript.

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Correspondence to Erhard Hohenester or Kevin P Campbell.

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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).

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