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

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

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...

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Bibliographic Details
Published in:Nature communications Vol. 14; no. 1; pp. 5237 - 15
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: London Nature Publishing Group UK 28-08-2023
Nature Publishing Group
Nature Portfolio
Subjects:
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49/22
49/23
631/92/458/1524
631/92/72/1202
639/638/549/974
639/638/92/221
639/638/92/605
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Animals
Arrays
Asparagine
Avidity
Azides
Bacteriophages
CD22 antigen
Complexity
DC-SIGN protein
Density
Gene Library
Genetic code
Glycan
Glycoconjugates
Glycosidases
Glycosylation
Heterogeneity
Humanities and Social Sciences
Ions
Lectins
Libraries
Mice
multidisciplinary
N-glycans
Organs
Phages
Polysaccharides
Protein interaction
Proteins
Science
Science (multidisciplinary)
Structural analysis
Synthesis
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Summary: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. Cellular glycosylation is complex and heterogeneous, which is challenging to reproduce synthetically. Here, the authors report on enzymatic remodelling of multivalent glycosylated bacteriophages to produce genetically encoded library of N -glycans which can be used to measure glycan-protein interactions with lectins on the surface of live cells and organs.
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ISSN:2041-1723
2041-1723
DOI:10.1038/s41467-023-40900-y