Shotgun ion mobility mass spectrometry sequencing of heparan sulfate saccharides

Miller, Rebecca L.; Guimond, Scott E.; Schwörer, Ralf; Zubkova, Olga V.; Tyler, Peter C.; Xu, Yongmei; Liu, Jian; Chopra, Pradeep; Boons, Geert-Jan; Grabarics, Márkó; Manz, Christian; Hofmann, Johanna; Karlsson, Niclas G.; Turnbull, Jeremy E.; Struwe, Weston B.; Pagel, Kevin

Abstract

Despite evident regulatory roles of heparan sulfate (HS) saccharides in numerous biological processes, definitive information on the bioactive sequences of these polymers is lacking, with only a handful of natural structures sequenced to date. Here, we develop a "Shotgun" Ion Mobility Mass Spectrometry Sequencing (SIMMS2) method in which intact HS saccharides are dissociated in an ion mobility mass spectrometer and collision cross section values of fragments measured. Matching of data for intact and fragment ions against known values for 36 fully defined HS saccharide structures (from di- to decasaccharides) permits unambiguous sequence determination of validated standards and unknown natural saccharides, notably including variants with 3O-sulfate groups. SIMMS2 analysis of two fibroblast growth factor-inhibiting hexasaccharides identified from a HS oligosaccharide library screen demonstrates that the approach allows elucidation of structure-activity relationships. SIMMS2 thus overcomes the bottleneck for decoding the informational content of functional HS motifs which is crucial for their future biomedical exploitation.

Citation

Miller, R. L., Guimond, S. E., Schwörer, R., Zubkova, O. V., Tyler, P. C., Xu, Y., …Pagel, K. (2020). Shotgun ion mobility mass spectrometry sequencing of heparan sulfate saccharides. Nature communications, 11, Article 1481. https://doi.org/10.1038/s41467-020-15284-y

• Journal Article Type: Article
• Acceptance Date: Feb 27, 2020
• Publication Date: Mar 20, 2020
• Journal: Nature Communications
• Print ISSN: 2041-1723
• Peer Reviewed: Peer Reviewed
• Volume: 11
• Article Number: 1481
• DOI: https://doi.org/10.1038/s41467-020-15284-y
• Keywords: Epitopes, Fibroblast Growth Factors, Glucuronic Acid, Heparin, Heparitin Sulfate, Ions, Mass Spectrometry, Oligosaccharides, Sequence Analysis, Structure-Activity Relationship, Sulfotransferases
• Publisher URL: http://doi.org/10.1038/s41467-020-15284-y