Dissecting structure-function of 3-O-sulfated heparin and engineered heparan sulfates
Karlsson, R.; Chopra, P.; Joshi, A.; Yang, Z.; Vakhrushev, S.Y.; Clausen, T.M.; Painter, C.D; Szekeres, G.P.; Chen, Y.-H.; Sandoval, D.R.; Hansen, L.; Esko, J.D.; Pagel, K.; Dyer, D.P.; Turnbull, J.E.; Clausen, H.; Boons, G.-J.; Miller, R.L. – 2021
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. The data provide new insights into 3-O-sulfate structure-function and demonstrate proof of concept for tailored cell-based synthesis of next-generation heparins.