Human Milk Oligosaccharide 2′-Fucosyllactose Inhibits Ligand Binding to C-Type Lectin DC-SIGN but Not to Langerin

Human milk oligosaccharides (HMOs) and its most abundant component, 2’-Fucosyllactose (2’-FL), are known to be immunomodulatory. Previously, it was shown that HMOs and 2’-FL bind to the C-type lectin receptor DC-SIGN. Here we show, using a ligand-receptor competition assay, that a whole mixture of HMOs from pooled human milk (HMOS) and 2’-FL inhibit the binding of the carbohydrate-binding receptor DC-SIGN to its prototypical ligands, fucose and the oligosaccharide Lewis-B, (Leb) in a dose-dependent way. Interestingly, such inhibition by HMOS and 2’-FL was not detected for another C-type lectin, Langerin, evolutionary similar to DC-SIGN. The cell-ligand competition assay using DC-SIGN expressing cells confirmed that 2’-FL inhibits the binding of DC-SIGN to Leb. Molecular dynamics (MD) simulations show that 2’-FL exists in a preorganized bioactive conformation before binding to DC-SIGN and this conformation is retained after binding to DC-SIGN. Leb has more flexible conformations and utilizes two binding modes, which operate one at a time via its two fucoses to bind to DC-SIGN. 2’-FL may have a reduced entropic penalty due to its preorganized state compared to Leb, and it has lower binding enthalpy, suggesting better binding to DC-SIGN. Thus, due to the better binding to DC-SIGN, 2’-FL may replace Leb from its binding pocket in DC-SIGN. MD simulations also showed that 2’-FL does not bind to Langerin. Our studies confirm 2’-FL as a specific ligand for DC-SIGN and suggest that 2’-FL can replace other DC-SIGN ligands from its binding pocket during ligand-receptor interactions in possible immunomodulatory processes.