Automated Glycan Assembly of (19) F-labeled Glycan Probes Enables High-Throughput NMR Studies of Protein-Glycan Interactions

利用自动化的(19)F标记糖探针进行糖链组装,可实现蛋白质-糖链相互作用的高通量核磁共振研究

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Abstract

Protein-glycan interactions mediate important biological processes, including pathogen host invasion and cellular communication. Herein, we showcase an expedite approach that integrates automated glycan assembly (AGA) of (19) F-labeled probes and high-throughput NMR methods, enabling the study of protein-glycan interactions. Synthetic Lewis type 2 antigens were screened against seven glycan binding proteins (GBPs), including DC-SIGN and BambL, respectively involved in HIV-1 and lung infections in immunocompromised patients, confirming the preference for fucosylated glycans (Le(x) , H type 2, Le(y) ). Previously unknown glycan-lectin weak interactions were detected, and thermodynamic data were obtained. Enzymatic reactions were monitored in real-time, delivering kinetic parameters. These results demonstrate the utility of AGA combined with (19) F NMR for the discovery and characterization of glycan-protein interactions, opening up new perspectives for (19) F-labeled complex glycans.

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