Ion mobility-tandem mass spectrometry of mucin-type O-glycans

Leïla Bechtella, Jin Chunsheng, Kerstin Fentker, Güney R. Ertürk, Marc Safferthal, Łukasz Polewski, Michael Götze, Simon Y. Graeber, Gaël M. Vos, Weston B. Struwe, Marcus A. Mall, Philipp Mertins, Niclas G. Karlsson and Kevin Pagel ()
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Leïla Bechtella: Freie Universität Berlin
Jin Chunsheng: University of Gothenburg
Kerstin Fentker: Freie Universität Berlin
Güney R. Ertürk: Freie Universität Berlin
Marc Safferthal: Freie Universität Berlin
Łukasz Polewski: Freie Universität Berlin
Michael Götze: Freie Universität Berlin
Simon Y. Graeber: corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin
Gaël M. Vos: Freie Universität Berlin
Weston B. Struwe: University of Oxford
Marcus A. Mall: corporate member of Freie Universität Berlin and Humboldt-Universität zu Berlin
Philipp Mertins: Max Delbrück Center for Molecular Medicine
Niclas G. Karlsson: University of Gothenburg
Kevin Pagel: Freie Universität Berlin

Nature Communications, 2024, vol. 15, issue 1, 1-10

Abstract: Abstract The dense O-glycosylation of mucins plays an important role in the defensive properties of the mucus hydrogel. Aberrant glycosylation is often correlated with inflammation and pathology such as COPD, cancer, and Crohn’s disease. The inherent complexity of glycans and the diversity in the O-core structure constitute fundamental challenges for the analysis of mucin-type O-glycans. Due to coexistence of multiple isomers, multidimensional workflows such as LC-MS are required. To separate the highly polar carbohydrates, porous graphitized carbon is often used as a stationary phase. However, LC-MS workflows are time-consuming and lack reproducibility. Here we present a rapid alternative for separating and identifying O-glycans released from mucins based on trapped ion mobility mass spectrometry. Compared to established LC-MS, the acquisition time is reduced from an hour to two minutes. To test the validity, the developed workflow was applied to sputum samples from cystic fibrosis patients to map O-glycosylation features associated with disease.

Date: 2024
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DOI: 10.1038/s41467-024-46825-4

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