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Shotgun ion mobility mass spectrometry sequencing of heparan sulfate saccharides

Artikel i vetenskaplig tidskrift
Författare R. L. Miller
S. E. Guimond
R. Schworer
O. V. Zubkova
P. C. Tyler
Y. M. Xu
J. Liu
P. Chopra
G. J. Boons
M. Grabarics
C. Manz
J. Hofmann
Niclas G. Karlsson
J. E. Turnbull
W. B. Struwe
K. Pagel
Publicerad i Nature Communications
Volym 11
Nummer/häfte 1
Sidor 12
ISSN 2041-1723
Publiceringsår 2020
Publicerad vid Institutionen för biomedicin
Sidor 12
Språk en
Länkar dx.doi.org/10.1038/s41467-020-15284...
Ämnesord electron detachment dissociation, collision cross-sections, glycosaminoglycan tetrasaccharides, chemoenzymatic synthesis, high-throughput, iduronic acid, oligosaccharides, identification, separation, conformation, Science & Technology - Other Topics
Ämneskategorier Strukturbiologi

Sammanfattning

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. Heparan sulfates (HS) contain functionally relevant structural motifs, but determining their monosaccharide sequence remains challenging. Here, the authors develop an ion mobility mass spectrometry-based method that allows unambiguous characterization of HS sequences and structure-activity relationships.

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