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

Autor: Bui DT; Department of Chemistry, University of Alberta, Edmonton T6G 2G2, Alberta, Canada., Favell J; Department of Chemistry, University of Alberta, Edmonton T6G 2G2, Alberta, Canada., Kitova EN; Department of Chemistry, University of Alberta, Edmonton T6G 2G2, Alberta, Canada., Li Z; Department of Chemistry, University of Alberta, Edmonton T6G 2G2, Alberta, Canada., McCord KA; Department of Chemistry, University of Alberta, Edmonton T6G 2G2, Alberta, Canada., Schmidt EN; Department of Chemistry, University of Alberta, Edmonton T6G 2G2, Alberta, Canada., Mozaneh F; Department of Chemistry, University of Alberta, Edmonton T6G 2G2, Alberta, Canada., Elaish M; Department of Cell Biology, University of Alberta, Edmonton T6G 2H7, AB, Canada.; Poultry Diseases Department, Faculty of Veterinary Medicine, Cairo University, Giza 12211, Egypt., El-Hawiet A; Department of Pharmacognosy, Faculty of Pharmacy, Alexandria University, Alexandria 21561, Egypt., St-Pierre Y; Institut National de la Recherche Scientifique (INRS), INRS-Centre Armand-Frappier Santé Biotechnologie, Laval H7 V 1B7, QC, Canada., Hobman TC; Department of Cell Biology, University of Alberta, Edmonton T6G 2H7, AB, Canada.; Department of Medical Microbiology and Immunology, University of Alberta, Edmonton T6G 2E1, AB, Canada.; Li Ka Shing Institute of Virology, University of Alberta, Edmonton T6G 2E1, Alberta, Canada., Macauley MS; Department of Chemistry, University of Alberta, Edmonton T6G 2G2, Alberta, Canada.; Department of Medical Microbiology and Immunology, University of Alberta, Edmonton T6G 2E1, AB, Canada., Mahal LK; Department of Chemistry, University of Alberta, Edmonton T6G 2G2, Alberta, Canada., Flynn MR; Department of Mechanical Engineering, Faculty of Engineering, University of Alberta, Edmonton T6G 1H9, Alberta, Canada., Klassen JS; Department of Chemistry, University of Alberta, Edmonton T6G 2G2, Alberta, Canada.
Jazyk: angličtina
Zdroj: ACS central science [ACS Cent Sci] 2023 Jun 15; Vol. 9 (7), pp. 1374-1387. Date of Electronic Publication: 2023 Jun 15 (Print Publication: 2023).
DOI: 10.1021/acscentsci.3c00294
Abstrakt: Native mass spectrometry (nMS) screening of natural glycan libraries against glycan-binding proteins (GBPs) is a powerful tool for ligand discovery. However, as the glycan concentrations are unknown, affinities cannot be measured directly from natural libraries. Here, we introduce Co ncentration- In dependent (COIN)-nMS, which enables quantitative screening of natural glycan libraries by exploiting slow mixing of solutions inside a nanoflow electrospray ionization emitter. The affinities ( K d ) of detected GBP-glycan interactions are determined, simultaneously, from nMS analysis of their time-dependent relative abundance changes. We establish the reliability of COIN-nMS using interactions between purified glycans and GBPs with known K d values. We also demonstrate the implementation of COIN-nMS using the catch-and-release (CaR)-nMS assay for glycosylated GBPs. The COIN-CaR-nMS results obtained for plant, fungal, viral, and human lectins with natural libraries containing hundreds of N -glycans and glycopeptides highlight the assay's versatility for discovering new ligands, precisely measuring their affinities, and uncovering "fine" specificities. Notably, the COIN-CaR-nMS results clarify the sialoglycan binding properties of the SARS-CoV-2 receptor binding domain and establish the recognition of monosialylated hybrid and biantennary N -glycans. Moreover, pharmacological depletion of host complex N -glycans reduces both pseudotyped virions and SARS-CoV-2 cell entry, suggesting that complex N -glycans may serve as attachment factors.
Competing Interests: The authors declare no competing financial interest.
(© 2023 The Authors. Published by American Chemical Society.)
Databáze: MEDLINE
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