A Novel Mechanism for Binding of Galactose-terminated Glycans by the C-type Carbohydrate Recognition Domain in Blood Dendritic Cell Antigen 2

Sabine A.F Jégouzo,Hadar Feinberg,Tabassum Dungarwalla,Kurt Drickamer,William I Weis,Maureen E Taylor

doi:10.1074/jbc.m115.660613

Sabine A.F Jégouzo, Hadar Feinberg + Show 4 more

Open Access

https://doi.org/10.1074/jbc.m115.660613

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Abstract

Blood dendritic cell antigen 2 (BDCA-2; also designated CLEC4C or CD303) is uniquely expressed on plasmacytoid dendritic cells. Stimulation of BDCA-2 with antibodies leads to an anti-inflammatory response in these cells, but the natural ligands for the receptor are not known. The C-type carbohydrate recognition domain in the extracellular portion of BDCA-2 contains a signature motif typical of C-type animal lectins that bind mannose, glucose, or GlcNAc, yet it has been reported that BDCA-2 binds selectively to galactose-terminated, biantennary N-linked glycans. A combination of glycan array analysis and binding competition studies with monosaccharides and natural and synthetic oligosaccharides have been used to define the binding epitope for BDCA-2 as the trisaccharide Galβ1-3/4GlcNAcβ1-2Man. X-ray crystallography and mutagenesis studies show that mannose is ligated to the conserved Ca(2+) in the primary binding site that is characteristic of C-type carbohydrate recognition domains, and the GlcNAc and galactose residues make additional interactions in a wide, shallow groove adjacent to the primary binding site. As predicted from these studies, BDCA-2 binds to IgG, which bears galactose-terminated glycans that are not commonly found attached to other serum glycoproteins. Thus, BDCA-2 has the potential to serve as a previously unrecognized immunoglobulin Fc receptor.

Highlights

Blood dendritic cell antigen 2 (BDCA-2) is an anti-inflammatory receptor uniquely expressed on plasmacytoid dendritic cells
Following expression of the CRD from BDCA-2 in E. coli as inclusion bodies, several protocols for renaturation were investigated, and attempts were made to purify the CRD on high density mannose-Sepharose resin, by analogy to protocols used for mincle and other C-type lectins
The protein did not stick tightly to the column and washed through in the presence of Ca2ϩ, the results provide evidence that BDCA-2 has at least weak mannose binding activity

Summary

Introduction

BDCA-2 is an anti-inflammatory receptor uniquely expressed on plasmacytoid dendritic cells. A combination of glycan array analysis and binding competition studies with monosaccharides and natural and synthetic oligosaccharides have been used to define the binding epitope for BDCA-2 as the trisaccharide Gal␤1–3/4GlcNAc␤1–2Man. X-ray crystallography and mutagenesis studies show that mannose is ligated to the conserved Ca2؉ in the primary binding site that is characteristic of C-type carbohydrate recognition domains, and the GlcNAc and galactose residues make additional interactions in a wide, shallow groove adjacent to the primary binding site. X-ray crystallography and mutagenesis studies show that mannose is ligated to the conserved Ca2؉ in the primary binding site that is characteristic of C-type carbohydrate recognition domains, and the GlcNAc and galactose residues make additional interactions in a wide, shallow groove adjacent to the primary binding site As predicted from these studies, BDCA-2 binds to IgG, which bears galactose-terminated glycans that are not commonly found attached to other serum glycoproteins

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Conclusion