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

• Published in: Science advances 2021-12, Vol.7 (52), p.eabl6026-eabl6026
• Main Authors: 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
• Format: Article
• Language: English
• Subjects: Biochemistry; Biomedicine and Life Sciences; Chemistry; SciAdv r-articles
• ISSN: 2375-2548; 2375-2548
• DOI: 10.1126/sciadv.abl6026

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- -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. The data provide new insights into 3- -sulfate structure-function and demonstrate proof of concept for tailored cell-based synthesis of next-generation heparins.