Dissecting structure-function of 3-O-sulfated heparin and engineered heparan sulfates

Karlsson, Richard; Chopra, Pradeep; Joshi, Apoorva; Yang, Zhang; Vakhrushev, Sergey Y.; Clausen, Thomas Mandel; Painter, Chelsea D.; Szekeres, Gergo P.; Chen, Yen-Hsi; Sandoval, Daniel R.; Hansen, Lars; Esko, Jeffrey D.; Pagel, Kevin; Dyer, Douglas P.; Turnbull, Jeremy E.; Clausen, Henrik; Boons, Geert-Jan; Miller, Rebecca L.

Abstract

Heparan sulfate (HS) polysaccharides are master regulators of diverse biological processes via sulfated motifs that can recruit specific proteins. 3-O-sulfation of HS/heparin is crucial for anticoagulant activity, but despite emerging evidence for roles in many other functions, a lack of tools for deciphering structure-function relationships has hampered advances. Here, we describe an approach integrating synthesis of 3-O-sulfated standards, comprehensive HS disaccharide profiling, and cell engineering to address this deficiency. Its application revealed previously unseen differences in 3-O-sulfated profiles of clinical heparins and 3-O-sulfotransferase (HS3ST)–specific variations in cell surface HS profiles. The latter correlated with functional differences in anticoagulant activity and binding to platelet factor 4 (PF4), which underlies heparin-induced thrombocytopenia, a known side effect of heparin. Unexpectedly, cells expressing the HS3ST4 isoenzyme generated HS with potent anticoagulant activity but weak PF4 binding.

Citation

Karlsson, R., Chopra, P., Joshi, A., Yang, Z., Vakhrushev, S. Y., Clausen, T. M., …Miller, R. L. (2021). Dissecting structure-function of 3-O-sulfated heparin and engineered heparan sulfates. Science Advances, 7(52), https://doi.org/10.1126/sciadv.abl6026

• Journal Article Type: Article
• Acceptance Date: Dec 22, 2021
• Online Publication Date: Dec 22, 2021
• Publication Date: Dec 24, 2021
• Deposit Date: May 30, 2023
• Journal: Science Advances
• Publisher: American Association for the Advancement of Science
• Peer Reviewed: Peer Reviewed
• Volume: 7
• Issue: 52
• DOI: https://doi.org/10.1126/sciadv.abl6026
• Keywords: Multidisciplinary