Chemoenzymatic Synthesis of 6‐Sulfo Sialyl Lewis x Containing Glycans to Probe the Receptor Specificity of MERS Coronavirus

ABSTRACT The initial attachment of Middle East Respiratory Syndrome Coronavirus (MERS-CoV) to host cell sialosides is critical for infection, yet its precise receptor specificity remains poorly understood. Here, we describe a chemoenzymatic methodology to synthesize a comprehensive panel of 6-sulfo sialyl Lewis x (6-sulfo-SLe x ) containing glycans. Our approach entails the enzymatic assembly of an oligo-lactosamine chain modified at specific positions with N -trifluoroacetyl-glucosamine (GlcNTFA) moieties. Mild base treatment removes the TFA group to yield glucosamine, which effectively blocks enzymatic fucosylation. By leveraging this approach alongside the unique substrate selectivity of GlcNAc-6-O-sulfotransferases 2 (CHST-2), we achieved the selective preparation of fucosylated 6-sulfo-SLe x glycans. Microarray screening of these printed glycans revealed that a 6-sulfo-SLe x derivative presented on an extended LacNAc chain is the preferred host receptor for MERS-CoV. Conjugation of this lead compound to a polyglycerol-based dendrimer generated a multivalent inhibitor that potently blocks hemagglutination of human red blood cells by the MERS-CoV spike protein N -terminal domain (NTD). Furthermore, computational modeling demonstrated that the fucose moiety does not directly contact the viral spike protein. Instead, it pre-organizes the ligand into a favorable conformation, enabling a critical salt bridge between the glycan’s sulfate group and the guanidinium side chain of viral residue Arg307.