Project description:Heparan sulfate (HS) mediates a wide range of protein binding interactions key to normal and pathological physiology. Though liquid chromatography coupled with mass spectrometry (LC-MS) based disaccharide composition analysis is able to profile changes in HS composition, the heterogeneity of modifications and the labile sulfate group present major challenges for liquid chromatography tandem mass spectrometry (LC-MS/MS) sequencing of the HS oligosaccharides that represent protein binding determinants. Here, we report online LC-MS/MS sequencing of HS oligosaccharides using hydrophilic interaction liquid chromatography (HILIC) and negative electron transfer dissociation (NETD). A series of synthetic HS oligosaccharides varying in chain length (tetramers and hexamers), number of sulfate groups (3-7), sulfate patterns (sulfate positional isomers), and uronic acid epimerization (epimers) were separated and sequenced. The LC elution order of isomeric compounds was associated with their fine structure. The application of an online cation exchange device (ion suppressor) enhanced the precursor charge states, and the subsequent NETD produced abundant glycosidic fragments, allowing the characterization of both lowly sulfated and highly sulfated HS oligosaccharides. Furthermore, the diagnostic cross-ring ions differentiated the 6-O sulfation and 3-O sulfation, allowing unambiguous structural assignment. Collectively, this LC-NETD-MS/MS method is a powerful tool for sequencing of heterogeneous HS mixtures and is applicable for the differentiation of both isomers and epimers, for the characterization of saccharide mixtures with a varying extent of sulfation and even for the determination of both predominant and rare modification motifs. Thus, LC-NETD-MS/MS has great potential for further application to biological studies.

| S-EPMC7161314 | biostudies-literature

Raw Metabolome (LC-MS) data of Hepatic Microtissues exposed to Acetaminophen (APAP)

Project description: Not available

| S-HECA159 | biostudies-other

Raw Metabolome (LC-MS) data of Hepatic Microtissues exposed to Phenytoin (PHE)

Project description: Not available

| S-HECA168 | biostudies-other

Raw Metabolome (LC-MS) data of Hepatic Microtissues not exposed (untreated; UNTR)

Project description: Not available

| S-HECA161 | biostudies-other

Raw Metabolome (LC-MS) data of Hepatic Microtissues exposed to DMSO (0.1%)

Project description: Not available