Custom Glycosylation of Cells and Proteins Using Cyclic Carbamate-Derivatized Oligosaccharides. Issue 11 (16th November 2017)

• Record Type: Journal Article
• Authors: Whitehead, Marek W.J.; Khanzhin, Nikolay; Borsig, Lubor; Hennet, Thierry
• Is Part Of: Cell chemical biology. Volume 24:Issue 11(2017)
• Page Start: 1336
• Page End: 1346.e3
• Publication Date: 2017-11-16
• Subjects: glycoconjugate -- glycoprotein -- lectin -- cell-surface engineering -- dendritic cell; Biochemistry -- Periodicals; 572.05
• DOI: 10.1016/j.chembiol.2017.08.012
• Languages: English
• ISSNs: 2451-9456

Abstract

Summary: The structural complexity of glycosylation restrains the functional characterization of glycans. We present a versatile carbohydrate ligation technique based on the reaction of cyclic carbamates with primary amines. Cyclic-carbamate-derivatized carbohydrates can be added to primary amine-containing molecules in aqueous solution to yield glycoconjugates. This method enabled the presentation of carbohydrate epitopes on live animal cells, as shown by the acquisition of E-selectin binding sites on mouse MC-38 cells decorated with 3-fucosyllactose or 3-fucosyl-3-sialyllactose. Ligation of 3- and 6-sialyllactose to Escherichia coli demonstrated the importance of sialic acid linkages in regulating complement factor H binding. Proteins were modified with oligosaccharides to study their role in stimulating cytokine secretion by dendritic cells, thus pointing to interactions between glycoproteins and phosphoinositide 3-kinase signaling in controlling interleukin-12, tumor necrosis factor alpha and interleukin-1β release. Overall, cyclic-carbamate-mediated ligation is useful to study the biology of carbohydrate epitopes on proteins and on cell membranes. Graphical Abstract: Highlights: Custom glycosylation of proteins and bacterial and animal cells Presentation of E-selectin ligands on cells by 3-fucosyl-3-sialyllactose coating Complement factor H binds α2-3 but not α2-6 linked sialic acid on coated E. coli Custom glycoproteins stimulate PI3K regulated inflammatory cytokine production Abstract : The structural heterogeneity impairs the functional characterization of glycan epitopes. Whitehead et al. use cyclic-carbamate-derivatized oligosaccharides to glycosylate proteins and bacterial and animal cells to examine the biological roles of such oligosaccharide epitopes.