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

Giulio Fittolani; Elena Shanina; Mónica Guberman; Peter H Seeberger; Christoph Rademacher; Martina Delbianco

doi:10.1002/anie.202102690

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

Angew Chem Int Ed Engl. 2021 Jun 7;60(24):13302-13309. doi: 10.1002/anie.202102690. Epub 2021 May 7.

Authors

Giulio Fittolani^{1

2}, Elena Shanina^{1

2}, Mónica Guberman^{1

3}, Peter H Seeberger^{1

2}, Christoph Rademacher^{1

2

4

5}, Martina Delbianco¹

Affiliations

¹ Department of Biomolecular Systems, Max Planck Institute of Colloids and Interfaces, Am Mühlenberg 1, 14476, Potsdam, Germany.
² Department of Chemistry and Biochemistry, Freie Universität Berlin, Arnimallee 22, 14195, Berlin, Germany.
³ Current address: Medicinal Chemistry, Leibniz-Forschungsinstitut für Molekulare Pharmakologie, Robert-Rössle Strasse 10, 13125, Berlin, Germany.
⁴ Current address: Department of Pharmaceutical Chemistry, University of Vienna, Althanstrasse 14, 1080, Vienna, Austria.
⁵ Current address: Department of Microbiology, Immunobiology and Genetics, Max F. Perutz Labs, Campus Vienna Biocenter 5, 1030, Vienna, Austria.

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.

Keywords: 19F NMR spectroscopy; automated glycan assembly; glycans; lectins; protein-glycan interactions.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Bacterial Proteins / chemistry
Bacterial Proteins / metabolism*
Binding Sites
Burkholderia / metabolism
Cell Adhesion Molecules / chemistry
Cell Adhesion Molecules / metabolism*
Fluorine / chemistry*
Glycosylation
Kinetics
Lectins / chemistry
Lectins / metabolism*
Lectins, C-Type / chemistry
Lectins, C-Type / metabolism*
Nuclear Magnetic Resonance, Biomolecular*
Polysaccharides / chemistry
Polysaccharides / metabolism*
Protein Binding
Receptors, Cell Surface / chemistry
Receptors, Cell Surface / metabolism*

Substances

Bacterial Proteins
Cell Adhesion Molecules
DC-specific ICAM-3 grabbing nonintegrin
Lectins
Lectins, C-Type
Polysaccharides
Receptors, Cell Surface
Fluorine