Harvey, DJ;
Scarff, CA;
Edgeworth, M;
Struwe, WB;
Pagel, K;
Thalassinos, K;
Crispin, M;
(2016)
Travelling-wave ion mobility and negative ion fragmentation of high-mannose N-glycans.
Journal of Mass Spectrometry
, 51
(3)
pp. 219-235.
10.1002/jms.3738.
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Abstract
The isomeric structure of high-mannose N-glycans can significantly impact biological recognition events. Here, the utility of travelling-wave ion mobility mass spectrometry for isomer separation of high-mannose N-glycans is investigated. Negative ion fragmentation using collision-induced dissociation gave more informative spectra than positive ion spectra with mass-different fragment ions characterizing many of the isomers. Isomer separation by ion mobility in both ionization modes was generally limited, with the arrival time distributions (ATD) often showing little sign of isomers. However, isomers could be partially resolved by plotting extracted fragment ATDs of the diagnostic fragment ions from the negative ion spectra, and the fragmentation spectra of the isomers could be extracted by using ions from limited areas of the ATD peak. In some cases, asymmetric ATDs were observed, but no isomers could be detected by fragmentation. In these cases, it was assumed that conformers or anomers were being separated. Collision cross sections of the isomers in positive and negative fragmentation mode were estimated from travelling-wave ion mobility mass spectrometry data using dextran glycans as calibrant. More complete collision cross section data were achieved in negative ion mode by utilizing the diagnostic fragment ions. Examples of isomer separations are shown for N-glycans released from the well-characterized glycoproteins chicken ovalbumin, porcine thyroglobulin and gp120 from the human immunodeficiency virus. In addition to the cross-sectional data, details of the negative ion collision-induced dissociation spectra of all resolved isomers are discussed.
| Type: | Article |
|---|---|
| Title: | Travelling-wave ion mobility and negative ion fragmentation of high-mannose N-glycans |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.1002/jms.3738 |
| Publisher version: | http://dx.doi.org/10.1002/jms.3738 |
| Language: | English |
| Additional information: | Copyright © 2016 John Wiley & Sons, Ltd. |
| Keywords: | Analytical, Spectroscopy, Biochemistry & Molecular Biology, Chemistry, T-wave ion mobility, N-linked glycosylation, isomers, high mannose, fragmentation, collision cross-sections, electron detachment dissociation, laser-desorption ionization, flight mass-spectrometry, linked glycans, structural-characterization, liquid-chromatography, isomeric oligosaccharides, saccharomyces-cerevisiae, h-1-nmr spectroscopy |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences > Structural and Molecular Biology |
| URI: | https://discovery.ucl.ac.uk/id/eprint/1481771 |
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