Integrated glycoproteomics identifies a role of N-glycosylation and galectin-1 on myogenesis and muscle development

Ronnie Blazev, Christopher Ashwood, Jodie L. Abrahams, Long H. Chung, Deanne Francis, Pengyi Yang, Kevin I. Watt, Hongwei Qian, Gregory A. Quaife-Ryan, James E. Hudson, Paul Gregorevic, Morten Thaysen-Andersen, Benjamin L. Parker*

*Corresponding author for this work

    Research output: Contribution to journalArticlepeer-review

    15 Citations (Scopus)
    37 Downloads (Pure)


    Many cell surface and secreted proteins are modified by the covalent addition of glycans that play an important role in the development of multicellular organisms. These glycan modifications enable communication between cells and the extracellular matrix via interactions with specific glycan-binding lectins and the regulation of receptor-mediated signaling. Aberrant protein glycosylation has been associated with the development of several muscular diseases, suggesting essential glycan- and lectin-mediated functions in myogenesis and muscle development, but our molecular understanding of the precise glycans, catalytic enzymes, and lectins involved remains only partially understood. Here, we quantified dynamic remodeling of the membrane-associated proteome during a time-course of myogenesis in cell culture. We observed wide-spread changes in the abundance of several important lectins and enzymes facilitating glycan biosynthesis. Glycomics-based quantification of released N-linked glycans confirmed remodeling of the glycome consistent with the regulation of glycosyltransferases and glycosidases responsible for their formation including a previously unknown digalactose-to-sialic acid switch supporting a functional role of these glycoepitopes in myogenesis. Furthermore, dynamic quantitative glycoproteomic analysis with multiplexed stable isotope labeling and analysis of enriched glycopeptides with multiple fragmentation approaches identified glycoproteins modified by these regulated glycans including several integrins and growth factor receptors. Myogenesis was also associated with the regulation of several lectins, most notably the upregulation of galectin-1 (LGALS1). CRISPR/Cas9-mediated deletion of Lgals1 inhibited differentiation and myotube formation, suggesting an early functional role of galectin-1 in the myogenic program. Importantly, similar changes in N-glycosylation and the upregulation of galectin-1 during postnatal skeletal muscle development were observed in mice. Treatment of new-born mice with recombinant adeno-associated viruses to overexpress galectin-1 in the musculature resulted in enhanced muscle mass. Our data form a valuable resource to further understand the glycobiology of myogenesis and will aid the development of intervention strategies to promote healthy muscle development or regeneration.

    Original languageEnglish
    Article number100030
    Pages (from-to)1-13
    Number of pages14
    JournalMolecular and Cellular Proteomics
    Publication statusPublished - 2021

    Bibliographical note

    Copyright the Author(s) 2021. Version archived for private and non-commercial use with the permission of the author/s and according to publisher conditions. For further rights please contact the publisher.


    • Glycoproteomics
    • glycomics
    • proteomics
    • glycosylation
    • myogenesis
    • myotubes
    • muscle development
    • galectin-1


    Dive into the research topics of 'Integrated glycoproteomics identifies a role of N-glycosylation and galectin-1 on myogenesis and muscle development'. Together they form a unique fingerprint.

    Cite this