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Structure and assembly of the dystrophin glycoprotein complex

Nature volume 637, pages 1252–1260 (2025)Cite this article

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Abstract

The dystrophin glycoprotein complex (DGC) has a crucial role in maintaining cell membrane stability and integrity by connecting the intracellular cytoskeleton with the surrounding extracellular matrix1,2,3. Dysfunction of dystrophin and its associated proteins results in muscular dystrophy, a disorder characterized by progressive muscle weakness and degeneration4,5. Despite the important roles of the DGC in physiology and pathology, its structural details remain largely unknown, hindering a comprehensive understanding of its assembly and function. Here we isolated the native DGC from mouse skeletal muscle and obtained its high-resolution structure. Our findings unveil a markedly divergent structure from the previous model of DGC assembly. Specifically, on the extracellular side, β-, γ- and δ-sarcoglycans co-fold to form a specialized, extracellular tower-like structure, which has a central role in complex assembly by providing binding sites for α-sarcoglycan and dystroglycan. In the transmembrane region, sarcoglycans and sarcospan flank and stabilize the single transmembrane helix of dystroglycan, rather than forming a subcomplex as previously proposed6,7,8. On the intracellular side, sarcoglycans and dystroglycan engage in assembly with the dystrophin–dystrobrevin subcomplex through extensive interaction with the ZZ domain of dystrophin. Collectively, these findings enhance our understanding of the structural linkage across the cell membrane and provide a foundation for the molecular interpretation of many muscular dystrophy-related mutations.

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Fig. 1: Cryo-EM reconstruction of the DGC from mouse skeletal muscle.
Fig. 2: Structure of the ECD tower.
Fig. 3: Assembly of the DGC in the extracellular and transmembrane regions.
Fig. 4: Interactions between dystrophin and DAPs.
Fig. 5: Structural mapping of muscular dystrophy-related mutations.

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Data availability

The cryo-EM maps of the mouse DGC have been deposited at the Electron Microscopy Data Bank (https://www.ebi.ac.uk/pdbe/emdb/) under the accession codes EMD-39569 and EMD-39568. The corresponding atomic coordinate data has been deposited at the Protein Data Bank (http://www.rcsb.org) under the accession code 8YT8. All data analysed during this study are included in this Article and its Supplementary Information. Any other relevant reagents and materials are available from the corresponding author upon request.

Code availability

No code was used for this study.

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Acknowledgements

The authors thank the cryo-EM Facility of Westlake University for providing support on cryo-EM data collection; Westlake University HPC Center for computational resources and related assistance; the Mass Spectrometry and Metabolomics Core Facility of Westlake University for mass spectrometry analysis. This work was supported by National Natural Science Foundation of China (32271261 to J.W. and 32271239 to Z.Y.), Zhejiang Provincial Natural Science Foundation of China (LR22C050003 to J.W.), Westlake University (1011103860222B1 to J.W. and 1011103560222B1 to Z.Y.) and Westlake Education Foundation (101486021901 to J.W. and 101456021901 to Z.Y.). Research reported in this publication was also supported by the National Institute of Neurological Disorders and Stroke of the National Institutes of Health under Award Number P50NS053672 to K.P.C. K.P.C is an investigator of the Howard Hughes Medical Institute.

Author information

Author notes

  1. These authors contributed equally: Li Wan, Xiaofei Ge, Qikui Xu, Gaoxingyu Huang

Authors and Affiliations

  1. Key Laboratory of Structural Biology of Zhejiang Province, School of Life Sciences, Westlake University, Hangzhou, China

    Li Wan, Qikui Xu, Gaoxingyu Huang, Zhen Yan & Jianping Wu

  2. Westlake Laboratory of Life Sciences and Biomedicine, Hangzhou, China

    Li Wan, Qikui Xu, Gaoxingyu Huang, Zhen Yan & Jianping Wu

  3. Institute of Biology, Westlake Institute for Advanced Study, Hangzhou, China

    Li Wan, Qikui Xu, Gaoxingyu Huang, Zhen Yan & Jianping Wu

  4. State Key Laboratory of Membrane Biology, Beijing Frontier Research Center for Biological Structure, School of Life Sciences, Tsinghua University, Beijing, China

    Xiaofei Ge

  5. Howard Hughes Medical Institute, University of Iowa Roy J. and Lucille A. Carver College of Medicine, Iowa City, IA, USA

    Tiandi Yang & Kevin P. Campbell

  6. Senator Paul D. Wellstone Muscular Dystrophy Specialized Research Center, University of Iowa Roy J. and Lucille A. Carver College of Medicine, Iowa City, IA, USA

    Tiandi Yang & Kevin P. Campbell

  7. Department of Molecular Physiology and Biophysics, University of Iowa Roy J. and Lucille A. Carver College of Medicine, Iowa City, IA, USA

    Tiandi Yang & Kevin P. Campbell

  8. Department of Neurology, University of Iowa Roy J. and Lucille A. Carver College of Medicine, Iowa City, IA, USA

    Tiandi Yang & Kevin P. Campbell

  9. Department of Immunology, Harvard Medical School, Boston, MA, USA

    Tiandi Yang

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Contributions

J.W. and Z.Y. conceived and supervised the project. J.W., Z.Y. and L.W. designed the experiments. L.W. prepared the protein samples, collected cryo-EM datasets and performed all other biochemical experiments. X.G., Q.X. and G.H. performed the cryo-EM data processing. Q.X. and J.W. built the atomic model. T.Y. and K.P.C. advised on DGC protein preparation and contributed to manuscript discussions. All authors contributed to data analysis. J.W., Z.Y. and L.W. wrote the manuscript with input from all co-authors.

Corresponding authors

Correspondence to Zhen Yan or Jianping Wu.

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The authors declare no competing interests.

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Nature thanks Jeffrey Chamberlain and the other, anonymous, reviewer(s) for their contribution to the peer review of this work. Peer review reports are available.

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Extended data figures and tables

Extended Data Fig. 1 Endogenous purification of the DGC from mouse skeletal muscle.

a, A classic schematic showing the overall organization of the DGC. The LG4 and LG5 domains of laminin-α2 interact with the glycans on α-DG. Inset: crystal structure of the LG4 and LG5 domains of laminin-α2 in complex with glycans (PDB: 5IK5). ECM: extracellular matrix; ABD: actin-binding domain; CR: cysteine-rich domain; CT: C-terminal domain. b, Size-exclusion chromatogram and corresponding SDS-PAGE analysis of the purified LG4 and LG5 domains of laminin-α2. Purifications were repeated independently at least three times with similar results. c, A diagram showing the purification procedure of the native DGC from mouse skeletal muscle. SEC: size exclusion chromatography; WB: western blot; MS: mass spectrometry. d, Size exclusion chromatography profile of the purified DGC sample. The shaded fractions were collected for cryo-EM and MS analysis. e, Western blot analysis of the gel filtration fractions against multiple DGC components. Each western blot was repeated at least twice with similar results. For gel source data, see Supplementary Fig. 1a. f, MS analysis of the purified DGC sample. Potential DGC components are listed in the order of decreasing peptide-spectrum match (PSM) scores. The DGC components observed in our model are highlighted in green. g, Peptide identification of dystrophin by MS analysis of the purified DGC sample. The identified regions are shaded in cyan, which account for a total of 67% sequence coverage.

Extended Data Fig. 2 Cryo-EM data analysis of the DGC.

a, A raw cryo-EM image of the purified DGC sample out of a dataset of 40,736 images. Representative particles are highlighted by green circles. Scale bar: 50 nm. b, Two-dimensional class averages. Box size: 469.6 Å. c, Gold-standard Fourier shell correlation (FSC) curves of the final maps. d, A flowchart of cryo-EM data processing. For details, see ‘Image processing’ in the Methods. Map 1 (EMDB-39569) is the map with the highest overall resolution and Map 2 (EMDB-39568) is the map with the clearest intracellular densities. Both maps were used to guide model building. The local resolutions of the two maps were estimated by cryoSPARC.

Extended Data Fig. 3 Density maps of the DGC components.

a, Density maps of selected segments of each DGC component. The density of dystrobrevin is presented in Fig. 4f. The boundaries of each segment and some bulky residues are labelled. b, The glycan densities of all identified glycosylation sites and the densities of three cholesterol-like lipids. The density maps were generated in ChimeraX.

Extended Data Fig. 4 Structural topology and sequence alignment of β-, γ-, and δ-sarcoglycans.

a, Topological diagram of β-, γ-, and δ-sarcoglycans. The β strands on face (a), (b), and (c) of the ECD tower are coloured in purple, green, and cyan, respectively. The anti-parallel β strands within the same face of the ECD tower are boxed. The β14 strand of β-sarcoglycan, which differs from the other two sarcoglycans, is highlighted by a red box. The β strands on each face of the ECD tower are connected by coloured dashed lines. b, Sequence alignment among β-, γ-, and δ-sarcoglycans. Secondary structure elements of β-sarcoglycan are labelled above the sequence and that of γ- and δ-sarcoglycans are labelled below the sequence. The N-terminal regions that are not modelled are indicated by dashed lines. Glycosylation sites are indicated by green boxes and asterisks. Disulfide bonds are labelled by orange lines. The UniProt IDs for each sequence are as follows: β-sarcoglycan: P82349, γ-sarcoglycan: P82348, δ-sarcoglycan: P82347. c, Structural comparison among several β-helical-containing proteins. The structures presented include the β-helical domains of the ECD tower of the DGC, VgrG (PDB: 6SK0), Pdp-VgrG (PDB: 6U9E), gp5 from the T4 bacteriophage (PDB: 1K28), and gpJ in closed/apo (PDB: 8XCK) and open/receptor-bound (PDB: 8XCJ) states. The relative rotational and overall height changes of gpJ between the apo and receptor-bound states are indicated by grey arrows and dashed lines, respectively.

Extended Data Fig. 5 Domain organization of dystroglycan and α-sarcoglycan.

a, Schematic diagrams of dystroglycan and α-sarcoglycan. The resolved extracellular domains of the two components are shaded in colour. Glycosylation sites and disulfide bonds are labelled. SP: signal peptide. b, Structural comparisons among the extracellular domains of dystroglycan and α-sarcoglycan. The red arrow indicates the reported dividing point between α-DG and β-DG, situated in the loop region between the β2 and β3 strands of the P domain. c, Structural overlay among the extracellular domains of dystroglycan and α-sarcoglycan. The three models are superimposed by their P domains. The double-headed arrow indicates conformational variations of the immunoglobulin-like domains.

Extended Data Fig. 6 Interaction between dystrophin and dystrobrevin.

a, Schematic diagram of dystrophin and dystrobrevin. b, Structure of CR domain of dystrophin fitted onto the cryo-EM map. The map is shown as a transparent surface. The red arrow indicates extra unassigned density near the zinc-binding site of the ZZ domain. c, Overall structure of the resolved domains of dystrophin and dystrobrevin. The crystal structure of human dystrophin in complex with the C-terminus of β-DG peptide (PDB:1EG4) is also presented for comparison. The domains are coloured in the same scheme as in a. d, Comparison of the crystal structure and the cryo-EM structure of the WW and EF-hand domains of dystrophin. The cryo-EM density map of this region is shown as a transparent surface. The red arrow indicates the conformational deviation of the WW domain between the two structures. S3059 and T3074 on the WW domain of dystrophin, and two tyrosine residues on the C-terminus of β-DG, which have been reported as phosphorylation sites, are shown as sticks. e, Structural overlay of the EF-hand domains in dystrobrevin and dystrophin. f, Comparison between the cryo-EM structure and the predicted structure by Alphafold3. The predicted structure between dystrophin (3049–3678) and dystrobrevin (1–746) suggests the presence of two major interaction interfaces. g, Gel filtration binding assay to verify the interaction between dystrobrevin and dystrophin. Left: dystrobrevin co-migrates with the GST-tagged dystrophin. Right: dystrobrevin does not co-migrate with GST alone. The co-migration of dystrobrevin and GST-tagged dystrophin is highlighted by a red box. The assay was repeated independently three times with similar results. For gel source data, see Supplementary Fig. 1b.

Extended Data Fig. 7 Recombinant expression of the extracellular domains of α-sarcoglycan and dystroglycan.

a, Size-exclusion chromatogram of wild-type and 19 disease-related mutations of α-sarcoglycan (1–250). In addition to the monomeric peak, the recombinantly expressed protein also exhibits an oligomeric peak, possibly due to heterologous overexpression in HEK293 cells. b, SDS-PAGE analysis of the purified wild-type and disease-related mutations of α-sarcoglycan. For gel source data, see Supplementary Fig. 1c. c, Size-exclusion chromatogram of wild-type and the C667F mutation of dystroglycan (492–712). d, SDS-PAGE analysis of the purified wild-type and the C667F mutation of dystroglycan. For gel source data, see Supplementary Fig. 1d.

Extended Data Fig. 8 A modified schematic of the DGC based on our structure.

In this model, sarcoglycans play a central role in complex assembly. In the extracellular side, β-, γ-, and δ-sarcoglycans co-fold to form a large ECD tower, which serves as docking sites for multiple extracellular domains from other components. In the transmembrane region, the sarcoglycans and sarcospan flank two sides of the transmembrane of β-DG, thereby stabilizing the latter. In the cytoplasmic region, sarcoglycans and β-DG directly interact with the ZZ domain of dystrophin. The structural features of the DGC, including the characteristic tilt angle of the ECD tower, enable it to efficiently connect the two sides of sarcolemma and transmit both longitudinal and lateral forces.

Extended Data Table 1 Statistics for data collection and structural refinement
Full size table
Extended Data Table 2 Summary of model building of the DGC
Full size table
Extended Data Table 3 Disease-related mutants on the DGC
Full size table

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Wan, L., Ge, X., Xu, Q. et al. Structure and assembly of the dystrophin glycoprotein complex. Nature 637, 1252–1260 (2025). https://doi.org/10.1038/s41586-024-08310-2

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