Chemoenzymatic Synthesis of Genetically-Encoded Multivalent Liquid N-glycan Arrays

Lin, C.-L.; Sojitra, M.; Carpenter, E. J.; Hayhoe, E. S.; Sarkar, S.; Volker, E. A.; Atrazhev, A.; Lowary, T. L.; Macauley, M. S.; Derda, R.

doi:10.1101/2022.08.05.503005

bioRxiv · 10.1101/2022.08.05.503005

Chemoenzymatic Synthesis of Genetically-Encoded Multivalent Liquid N-glycan Arrays

Lin, C.-L.·Sojitra, M.·Carpenter, E. J.·Hayhoe, E. S.·Sarkar, S.·Volker, E. A.·Atrazhev, A.·Lowary, T. L.·Macauley, M. S.·Derda, R.

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Abstract

A hallmark of cellular glycosylation is its chemical complexity and heterogeneity, which can be challenging to capture synthetically. Using chemoenzymatic synthesis on M13 phage, we produce a genetically-encoded liquid glycan array (LiGA) of biantennary complex type N-glycans. Ligation of azido-functionalized sialylglycosyl-asparagine derived from egg yolk to phage functionalized with 50-1000 copies of dibenzocyclooctyne produced divergent intermediate that can be trimmed by glycosidases and extended by glycosyltransferases to yield a library of phages with different N-glycans. Post-reaction analysis by MALDI-TOF MS provided a rigorous approach to confirm N-glycan structure and density, both of which were encoded in the bacteriophage DNA. The binding of this N-glycan LiGA by ten lectins, including CD22 or DC-SIGN expressed on live cells, uncovered an optimal structure/density combination for recognition. Injection of the LiGA into mice identified glycoconjugates with structures and avidity necessary for enrichment in specific organs. This work provides an unprecedented quantitative evaluation of the interaction of complex N-glycans with GBPs in vitro and in vivo.

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Lin, C.-L., Sojitra, M., Carpenter, E. J., Hayhoe, E. S., Sarkar, S., Volker, E. A., Atrazhev, A., Lowary, T. L., Macauley, M. S., Derda, R.. 2022-08-06. Chemoenzymatic Synthesis of Genetically-Encoded Multivalent Liquid N-glycan Arrays. https://doi.org/10.1101/2022.08.05.503005

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