Chemoenzymatic synthesis of genetically-encoded multivalent liquid N-glycan arrays

• Published in: Nature communications Vol. 14; no. 1; p. 5237
• Main Authors: Lin, Chih-Lan, Sojitra, Mirat, Carpenter, Eric J, Hayhoe, Ellen S, Sarkar, Susmita, Volker, Elizabeth A, Wang, Chao, Bui, Duong T, Yang, Loretta, Klassen, John S, Wu, Peng, Macauley, Matthew S, Lowary, Todd L, Derda, Ratmir
• Format: Journal Article
• Language: English
• Published: England Nature Publishing Group 28.08.2023; Nature Publishing Group UK; Nature Portfolio
• Subjects: Animals; Arrays; Asparagine; Avidity; Azides; Bacteriophages; CD22 antigen; Cell surface; Complexity; DC-SIGN protein; Density; Deoxyribonucleic acid; DNA; Gene Library; Genetic code; Glycan; Glycoconjugates; Glycosidases; Glycosylation; Heterogeneity; Ions; Lectins; Libraries; Mice; N-glycans; Organs; Phages; Polysaccharides; Protein interaction; Proteins; Structural analysis; Synthesis
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-023-40900-y

Cellular glycosylation is characterized by chemical complexity and heterogeneity, which is challenging to reproduce synthetically. Here we show chemoenzymatic synthesis on phage to produce a genetically-encoded liquid glycan array (LiGA) of complex type N-glycans. Implementing the approach involved by ligating an azide-containing sialylglycosyl-asparagine to phage functionalized with 50-1000 copies of dibenzocyclooctyne. The resulting intermediate can be trimmed by glycosidases and extended by glycosyltransferases yielding a phage library with different N-glycans. Post-reaction analysis by MALDI-TOF MS allows rigorous characterization of N-glycan structure and mean density, which are both encoded in the phage DNA. Use of this LiGA with fifteen glycan-binding proteins, including CD22 or DC-SIGN on cells, reveals optimal structure/density combinations for recognition. Injection of the LiGA into mice identifies glycoconjugates with structures and avidity necessary for enrichment in specific organs. This work provides a quantitative evaluation of the interaction of complex N-glycans with GBPs in vitro and in vivo.