Send to

Choose Destination
Commun Biol. 2019 Jul 22;2:268. doi: 10.1038/s42003-019-0507-2. eCollection 2019.

A quantitative, high-throughput method identifies protein-glycan interactions via mass spectrometry.

Author information

1Alberta Glycomics Centre and Department of Chemistry, University of Alberta, Edmonton, AB T6G 2G2 Canada.
2Department of Medical Microbiology and Immunology, University of Alberta, Edmonton, AB T6G 2E1 Canada.


Glycan binding by glycan-binding proteins and processing by carbohydrate-active enzymes is implicated in physiological and pathophysiological processes. Comprehensive mapping of glycan interactions is essential to understanding of glycan-mediated biology and can guide the development of new diagnostics and therapeutics. Here, we introduce the competitive universal proxy receptor assay (CUPRA), which combines electrospray ionization mass spectrometry, competitive binding and heterobifunctional glycan-based ligands to give a quantitative high-throughput method for screening glycan libraries against glycan-binding and glycan-processing proteins. Application of the assay to human (siglec-2), plant (Sambucus nigra and Maackia amurensis lectins) and bacterial (cholera toxin, and family 51 carbohydrate binding module) proteins allowed for the identification of ligands with affinities (K d) ≤ 1 mM. The assay is unprecedentedly versatile and can be applied to natural libraries and, when implemented in a time-resolved manner, provides a quantitative measure of the activities and substrate specificity of carbohydrate-active enzymes.


High-throughput screening; Mass spectrometry

Conflict of interest statement

Competing interestsThe authors declare no competing interests.

Supplemental Content

Full text links

Icon for PubMed Central
Loading ...
Support Center