Systematic synthesis of bisected N-glycans and unique recognitions by glycan-binding proteins

Xuefeng Cao; Shuaishuai Wang; Madhusudhan Reddy Gadi; Ding Liu; Peng G. Wang; Xiu-Feng Wan; Jian Zhang; Xi Chen; Lauren E. Pepi; Parastoo Azadi; Lei Li

doi:10.1039/D1SC05435J

Systematic synthesis of bisected N-glycans and unique recognitions by glycan-binding proteins†

Xuefeng Cao,‡^a Shuaishuai Wang,

‡^a Madhusudhan Reddy Gadi,

‡^a Ding Liu,^a Peng G. Wang,^a Xiu-Feng Wan,^bcde Jian Zhang,^f Xi Chen,

^g Lauren E. Pepi,^h Parastoo Azadi^h and Lei Li

*^a

Author affiliations

* Corresponding authors

^a Department of Chemistry, Georgia State University, Atlanta, GA, USA
E-mail: lli22@gsu.edu

^b MU Center for Research on Influenza Systems Biology (CRISB), University of Missouri, Columbia, MO, USA

^c Department of Molecular Microbiology and Immunology, School of Medicine, University of Missouri, Columbia, MO, USA

^d Bond Life Sciences Center, University of Missouri, Columbia, MO, USA

^e Department of Electrical Engineering & Computer Science, College of Engineering, University of Missouri, Columbia, MO, USA

^f Z Biotech, LLC, Aurora, CO, USA

^g Department of Chemistry, University of California, One Shields Avenue, Davis, CA, USA

^h Complex Carbohydrate Research Center, University of Georgia, Athens, GA, USA

Abstract

Bisected N-glycans represent a unique class of protein N-glycans that play critical roles in many biological processes. Herein, we describe the systematic synthesis of these structures. A bisected N-glycan hexasaccharide was chemically assembled with two orthogonal protecting groups attached at the C2 of the branching mannose residues, followed by sequential installation of GlcNAc and LacNAc building blocks to afford two asymmetric bisecting “cores”. Subsequent enzymatic modular extension of the “cores” yielded a comprehensive library of biantennary N-glycans containing the bisecting GlcNAc and presenting 6 common glycan determinants in a combinatorial fashion. These bisected N-glycans and their non-bisected counterparts were used to construct a distinctive glycan microarray to study their recognition by a wide variety of glycan-binding proteins (GBPs), including plant lectins, animal lectins, and influenza A virus hemagglutinins. Significantly, the bisecting GlcNAc could bestow (PHA-L, rDCIR2), enhance (PHA-E), or abolish (ConA, GNL, anti-CD15s antibody, etc.) N-glycan recognition of specific GBPs, and is tolerated by many others. In summary, synthesized compounds and the unique glycan microarray provide ideal standards and tools for glycoanalysis and functional glycomic studies. The microarray data provide new information regarding the fine details of N-glycan recognition by GBPs, and in turn improve their applications.

Chemical Science

Systematic synthesis of bisected N-glycans and unique recognitions by glycan-binding proteins†

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Systematic synthesis of bisected N-glycans and unique recognitions by glycan-binding proteins

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