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Absolute Affinities from Quantitative Shotgun Glycomics Using Concentration-Independent (COIN) Native Mass Spectrometry


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Bui, Duong T.; Favell, James; Kitova, Elena N.; Li, Zhixiong; McCord, Kelli A.; Schmidt, Edward N.; Mozaneh, Fahima; Elaish, Mohamed; El-Hawiet, Amr; St-Pierre, Yves ORCID logoORCID: https://orcid.org/0000-0002-1948-2041; Hobman, Tom C.; Macauley, Matthew S.; Mahal, Lara K.; Flynn, Morris R. et Klassen, John S. (2023). Absolute Affinities from Quantitative Shotgun Glycomics Using Concentration-Independent (COIN) Native Mass Spectrometry ACS Central Science , vol. 9 , nº 7. pp. 1374-1387. DOI: 10.1021/acscentsci.3c00294.

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We describea breakthrough screening technology that enablesthe measurement of absolute ligand affinities for glycan-binding proteinsusing natural libraries of glycans at unknown concentrations Native mass spectrometry (nMS) screening of natural glycanlibrariesagainst glycan-binding proteins (GBPs) is a powerful tool for liganddiscovery. However, as the glycan concentrations are unknown, affinitiescannot be measured directly from natural libraries. Here, we introduce Concentration-Independent (COIN)-nMS, which enablesquantitative screening of natural glycan libraries by exploiting slowmixing of solutions inside a nanoflow electrospray ionization emitter.The affinities (K (d)) of detected GBP-glycaninteractions are determined, simultaneously, from nMS analysis oftheir time-dependent relative abundance changes. We establish thereliability of COIN-nMS using interactions between purified glycansand GBPs with known K (d) values. We alsodemonstrate the implementation of COIN-nMS using the catch-and-release(CaR)-nMS assay for glycosylated GBPs. The COIN-CaR-nMS results obtainedfor plant, fungal, viral, and human lectins with natural librariescontaining hundreds of N-glycans and glycopeptideshighlight the assay's versatility for discovering new ligands,precisely measuring their affinities, and uncovering "fine"specificities. Notably, the COIN-CaR-nMS results clarify the sialoglycanbinding properties of the SARS-CoV-2 receptor binding domain and establishthe recognition of monosialylated hybrid and biantennary N-glycans. Moreover, pharmacological depletion of host complex N-glycans reduces both pseudotyped virions and SARS-CoV-2cell entry, suggesting that complex N-glycans mayserve as attachment factors.

Type de document: Article
Mots-clés libres: -
Centre: Centre INRS-Institut Armand Frappier
Date de dépôt: 11 déc. 2023 15:19
Dernière modification: 11 déc. 2023 15:19
URI: https://espace.inrs.ca/id/eprint/13629

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