An Atlas of Human Glycosylation Pathways Enables Display of the Human Glycome by Gene Engineered Cells

• Published in: Molecular cell Vol. 75; no. 2; pp. 394 - 407.e5
• Main Authors: Narimatsu, Yoshiki, Joshi, Hiren J., Nason, Rebecca, Van Coillie, Julie, Karlsson, Richard, Sun, Lingbo, Ye, Zilu, Chen, Yen-Hsi, Schjoldager, Katrine T., Steentoft, Catharina, Furukawa, Sanae, Bensing, Barbara A., Sullam, Paul M., Thompson, Andrew J., Paulson, James C., Büll, Christian, Adema, Gosse J., Mandel, Ulla, Hansen, Lars, Bennett, Eric Paul, Varki, Ajit, Vakhrushev, Sergey Y., Yang, Zhang, Clausen, Henrik
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 25.07.2019
• Subjects: adhesin; adhesins; biosynthesis; carbohydrate; Epitopes - genetics; Epitopes - immunology; galectin; genes; Genetic Engineering; glycan array; glycoconjugates; glycoengineering; Glycosylation; glycosyltransferase; Glycosyltransferases - genetics; haptens; HEK293 Cells; human cell lines; Humans; lectin; Metabolic Networks and Pathways - genetics; microarray; oligosaccharides; Oligosaccharides - genetics; polysaccharides; Polysaccharides - chemistry; Polysaccharides - classification; Polysaccharides - genetics; Polysaccharides - immunology; Proteins - genetics; Proteins - immunology; siglec; transferases; microarray; adhesin; glycoengineering; glycosyltransferase; lectin; galectin; glycosylation; glycan array; siglec; carbohydrate
• ISSN: 1097-2765; 1097-4164; 1097-4164
• DOI: 10.1016/j.molcel.2019.05.017

The structural diversity of glycans on cells—the glycome—is vast and complex to decipher. Glycan arrays display oligosaccharides and are used to report glycan hapten binding epitopes. Glycan arrays are limited resources and present saccharides without the context of other glycans and glycoconjugates. We used maps of glycosylation pathways to generate a library of isogenic HEK293 cells with combinatorially engineered glycosylation capacities designed to display and dissect the genetic, biosynthetic, and structural basis for glycan binding in a natural context. The cell-based glycan array is self-renewable and reports glycosyltransferase genes required (or blocking) for interactions through logical sequential biosynthetic steps, which is predictive of structural glycan features involved and provides instructions for synthesis, recombinant production, and genetic dissection strategies. Broad utility of the cell-based glycan array is demonstrated, and we uncover higher order binding of microbial adhesins to clustered patches of O-glycans organized by their presentation on proteins. [Display omitted] •Human glycosyltransferases (170 GTf genes) organized in glycosylation pathway maps•The human glycome displayed in a natural context on the cell surface•Sustainable cell-based array resource to dissect biological functions of glycans•Microbial adhesins may bind to clustered patches of O-glycans Narimatsu et al. display the diversity of human sugars on the surface of a library of cells by genetically engineering the cellular glycosylation machinery. Sugars on the cell surface play important roles in interactions with the environment, and the cell library developed opens for studies of biological interactions with sugars.