Tandem Mass Spectrometry for Structural Characterization of Doubly-Charged N-Linked Glycopeptides.

H-T Kitty Wong, Xiangfeng Chen, Simin Zhang, T-Y Lui, D Hu, T-W Dominic Chan

Journal of the American Society for Mass Spectrometry 2022 Aug 03

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Three dissociation methods, including collision-induced dissociation (CID), electron capture dissociation (ECD), and electronic excitation dissociation (EED), were systematically compared for structural characterization of doubly charged glycopeptide. CID produced distinctively different tandem mass spectra for glycopeptide adducted with different charge carriers. Protonated species produced mainly glycosidic cleavages in high abundance. CID of magnesiated glycopeptide formed more cross-ring cleavages, whereas doubly sodiated species produced cleavages at both glycan and peptide moieties. The effect of charge carriers on the fragmentation in ECD and EED was lower than that in CID. ECD produced mainly peptide backbone cleavages but limited cleavages at the glycan moiety, whereas EED of glycopeptide resulted in extensive fragmentation throughout the molecular ion regardless of the charge carriers. Magnesiated species gave, however, more cross-ring cleavages than other charge carriers did. These results demonstrated that EED of magnesiated species could be used as a one-step dissociation method for comprehensive structural analysis of glycopeptides.

Citation

H-T Kitty Wong, Xiangfeng Chen, Simin Zhang, T-Y Lui, D Hu, T-W Dominic Chan. Tandem Mass Spectrometry for Structural Characterization of Doubly-Charged N-Linked Glycopeptides. Journal of the American Society for Mass Spectrometry. 2022 Aug 03;33(8):1458-1464