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2013 ; 85
(12
): 5787-95
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An approach for separation and complete structural sequencing of heparin/heparan
sulfate-like oligosaccharides
#MMPMID23659663
Huang R
; Liu J
; Sharp JS
Anal Chem
2013[Jun]; 85
(12
): 5787-95
PMID23659663
show ga
As members of the glycosaminoglycan (GAG) family, heparin and heparan sulfate
(HS) are responsible for mediation of a wide range of essential biological
actions, most of which are mediated by specific patterns of modifications of
regions of these polysaccharides. To fully understand the regulation of HS
modification and the biological function of HS through its interactions with
protein ligands, it is essential to know the specific HS sequences present.
However, the sequencing of mixtures of HS oligosaccharides presents major
challenges due to the lability of the sulfate modifications, as well as
difficulties in separating isomeric HS chains. Here, we apply a sequential
chemical derivatization strategy involving permethylation, desulfation, and
trideuteroperacetylation to label original sulfation sites with stable and
hydrophobic trideuteroacetyl groups. The derivatization chemistry differentiates
between all possible heparin/HS sequences solely by glycosidic bond cleavages,
without the need to generate cross-ring cleavages. This derivatization strategy
combined with LC-MS/MS analysis has been used to separate and sequence five
synthetic HS-like oligosaccharides of sizes up to dodecasaccharide, as well as a
highly sulfated Arixtra-like heptamer. This strategy offers a unique capability
for the sequencing of microgram quantities of HS oligosaccharide mixtures by
LC-MS/MS.