Structural basis for human chondroitin sulfate chain polymerization
Chondroitin sulfates are complex polysaccharide chains that regulate various biological processes at the cell surface and within the extracellular matrix. Here, we identify four heterodimeric complexes responsible for chondroitin sulfate chain polymerization in humans: CHSY1-CHPF, CHSY1-CHPF2, CHSY3-CHPF, and CHSY3-CHPF2. Using a custom-tailored in vitro glycosylation assay based on chemo-enzymatically synthesized fluorescent substrates, we demonstrate that all four complexes exhibit chain polymerization activity. The cryo-EM structure of the CHSY3-CHPF complex provides molecular insights into the chondroitin sulfate chain polymerization reaction. The architecture of the catalytic sites suggests that CHSY1 and CHSY3 are enzymatically active, while CHPF and CHPF2 primarily play a stabilizing role. Mutational analysis of purified enzyme complexes, combined with an in cellulo complementation assay, confirms that only CHSY1 and CHSY3 have bifunctional glycosyltransferase activities. Based on the spatial arrangement of the catalytic sites, we propose that chondroitin sulfate chain polymerization follows a non-processive, distributive mechanism.
Authors
- Mélanie Friedel-Arboleas
- R.L. Cordeiro (ORCID: https://orcid.org/0000-0002-8827-8116)
- Elisabetta Boeri Erba (ORCID: https://orcid.org/0000-0002-5910-4707)
- Rebekka Wild (ORCID: https://orcid.org/0000-0003-2025-7228)
- Huazhang Shu (ORCID: https://orcid.org/0009-0005-8911-952X)
- Margaux Molinas (ORCID: https://orcid.org/0000-0002-8795-0425)
- Sylvain D. Vallet (ORCID: https://orcid.org/0000-0001-9885-7499)
- Rebecca L. Miller (ORCID: https://orcid.org/0000-0001-8574-1948)
- Magnus Nørregaard Grønset (ORCID: https://orcid.org/0000-0002-2582-5063)
- Marie Bourgeais (ORCID: https://orcid.org/0009-0002-8572-2362)
- Margot Weber (ORCID: https://orcid.org/0000-0002-8540-1924)
- Pradip Kumar Dutta (ORCID: https://orcid.org/0009-0006-2435-020X)
Institutions
- University of Copenhagen (DK)
- Centre National de la Recherche Scientifique (FR)
- Commissariat à l'Énergie Atomique et aux Énergies Alternatives (FR)
- CEA Grenoble (FR)
- Institut de Biologie Structurale (FR)
- Université Grenoble Alpes (FR)
Publication Details
- Journal
- Nature Communications
- Published
- 2025-11-26
- DOI
- https://doi.org/10.1038/s41467-025-66787-5
- Primary Topic
- Proteoglycans and glycosaminoglycans research
- Type
- article
- Field-Weighted Citation Impact
- 0.00
Funders
- Infrastructures en Biologie Santé et Agronomie
- Equipex
- Agence Nationale de la Recherche
- European Synchrotron Radiation Facility
- Danmarks Grundforskningsfond
- Fondation pour la Recherche Médicale
- Novo Nordisk
- Centre National de la Recherche Scientifique
- Fondation Bettencourt Schueller
- Novo Nordisk Fonden
- Région Auvergne-Rhône-Alpes
- French Infrastructure for Integrated Structural Biology