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Discrimination of Isomers of Released N- and O-Glycans Using Diagnostic Product Ions in Negative Ion PGC-LC-ESI-MS/MS

ABSTRACT: We used porous graphitized carbon-LC-ESI-MS/MS to separate and detect released N- and O-glycan isomers from mammalian model glycoproteins using negative mode resonance activation CID-MS/MS. By interrogating similar fragment spectra from closely related glycan isomers that differ only in arm position and sialyl linkage, product fragment ions for discrimination between these features were discovered. These diagnostic ions were shown to be useful for isomer discrimination using both linear and 3D ion trap mass spectrometers when analyzing complex glycan mixtures from cell lysates. This platform-independent workflow can potentially be extended to automate the characterization and quantitation of other challenging glycan isomers.

INSTRUMENT(S): Velos Plus

ORGANISM(S): Bos Taurus (ncbitaxon:9913) Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Nicolle H. Packer

PROVIDER: MSV000083688 | MassIVE | Mon Apr 15 13:40:00 BST 2019

REPOSITORIES: MassIVE

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Discrimination of Isomers of Released N- and O-Glycans Using Diagnostic Product Ions in Negative Ion PGC-LC-ESI-MS/MS.

Ashwood Christopher C Lin Chi-Hung CH Thaysen-Andersen Morten M Packer Nicolle H NH

Journal of the American Society for Mass Spectrometry 20180330 6

Profiling cellular protein glycosylation is challenging due to the presence of highly similar glycan structures that play diverse roles in cellular physiology. As the anomericity and the exact linkage type of a single glycosidic bond can influence glycan function, there is a demand for improved and automated methods to confirm detailed structural features and to discriminate between structurally similar isomers, overcoming a significant bottleneck in the analysis of data generated by glycomics e ...[more]

PMID: 29603058

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