Profiling of glycosphingolipids with SCDase digestion and HPLC-FLD-MS

Chakraberty, Radhika; Reiz, Béla; Cairo, Christopher W.

doi:10.1016/j.ab.2021.114361

Cited by 2 publications

(2 citation statements)

References 67 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Although multiple groups have reported the characterization and quantification of GM3 and GD3 in bovine milk, these reports all listed the total composition of the total ceramide rather than individual values for the carbon numbers and degree(s) of unsaturation of the LCB and FA. ,,, A limited number of studies have determined the range of LCB and FA groups present in bovine milk. However, these values were obtained by first releasing the glycan headgroup and then analyzing the fatty acyl and LCB moieties separately. ,, As noted above, analysis of the intact molecules should be more informative because it provides the means to obtain the most biologically relevant information about the combinations of LCB and FA groups present in a given sample. The UVPD MS/MS fragmentation of a few abundant molecular species of GM3 derived from bovine milk has been reported, but, to the best of our knowledge, there has not been a comprehensive analysis, by tandem MS or other analytical methods, of the diversity of ceramide compositions (in terms of LCB and FA) present in the individual intact molecular species of bovine or human milk gangliosides.…”

Section: Resultsmentioning

confidence: 99%

“…However, these values were obtained by first releasing the glycan headgroup and then analyzing the fatty acyl and LCB moieties separately. 23,26,27 As noted above, analysis of the intact molecules should be more informative because it provides the means to obtain the most biologically relevant information about the combinations of LCB and FA groups present in a given sample. The UVPD MS/ MS fragmentation of a few abundant molecular species of GM3 derived from bovine milk has been reported, 22 but, to the best of our knowledge, there has not been a comprehensive analysis, by tandem MS or other analytical methods, of the diversity of ceramide compositions (in terms of LCB and FA) present in the individual intact molecular species of bovine or human milk gangliosides.…”

Section: Workflow For the Characterization Of Ceramide Compositions O...mentioning

confidence: 99%

See 1 more Smart Citation

Defining the Ceramide Composition of Bovine and Human Milk Gangliosides by Direct Infusion ESI-CID Tandem Mass Spectrometry of Native and Permethylated Molecular Species

Liyanage,

Xia,

Ringler

et al. 2023

Anal. Chem.

View full text Add to dashboard Cite

Gangliosides are glycosphingolipids composed of an oligosaccharide that contains one or more sialic acid residues and is linked to a ceramide, a lipid composed of a long chain base (LCB) that bears an amide-linked fatty acyl group (FA). The ceramide portions of gangliosides are embedded in cell membranes; the exposed glycans interact with the extracellular environment. Gangliosides play a myriad of roles in activities such as cell–cell communication, formation of lipid rafts, cellular adhesion, calcium homeostasis, host-pathogen interaction, and viral invasion. Although the epitopes responsible for the interactions of gangliosides are located in the glycan, the epitope presentation is strongly influenced by the orientation of the attached ceramide within the lipid membrane, a feature that depends on the details of its structure, that is, the specific LCB and FA. Since the identities of both the glycan and the ceramide affect the activity of gangliosides, it is important to characterize the individual intact molecular forms. We report here a mass spectrometry-based method that combines the information gained from low-energy collision-induced dissociation (CID) measurements for the determination of the glycan with tandem mass spectra obtained at stepped higher-energy CID for the detailed characterization of the LCB and FA components of intact gangliosides. We provide results from applications of this method to the analysis of gangliosides present in bovine and human milk in order to demonstrate the assignment of LCB and FA for intact gangliosides and differential detection of isomeric ceramide structures.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Workflow For the Characterization Of Ceramide Compositions O...mentioning

confidence: 99%

Defining the Ceramide Composition of Bovine and Human Milk Gangliosides by Direct Infusion ESI-CID Tandem Mass Spectrometry of Native and Permethylated Molecular Species

Liyanage,

Xia,

Ringler

et al. 2023

Anal. Chem.

View full text Add to dashboard Cite

show abstract

Fucosylated glycoproteins and fucosylated glycolipids play opposing roles in cholera intoxication

Ghorashi

Boucher

Archer-Hartmann³

et al. 2023

Preprint

View full text Add to dashboard Cite

SUMMARYCholera toxin (CT) is the etiological agent of cholera. Here we report that multiple classes of fucosylated glycoconjugates function in CT binding and intoxication of intestinal epithelial cells. In Colo205 cells, knockout of B3GNT5, the enzyme required for synthesis of lacto- and neolacto-series glycosphingolipids (GSLs), reduces CT binding but sensitizes cells to intoxication. Overexpressing B3GNT5 to generate more fucosylated GSLs confers protection against intoxication, indicating that fucosylated GSLs act as decoy receptors for CT. Knockout (KO) of B3GALT5 causes increased production of fucosylated O-linked and N-linked glycoproteins, and leads to increased CT binding and intoxication. Knockout of B3GNT5 in B3GALT5 KO cells eliminates production of fucosylated GSLs but increases intoxication, identifying fucosylated glycoproteins as functional receptors for CT. These findings provide insight into molecular determinants regulating CT sensitivity of host cells.

show abstract

Profiling of glycosphingolipids with SCDase digestion and HPLC-FLD-MS

Cited by 2 publications

References 67 publications

Defining the Ceramide Composition of Bovine and Human Milk Gangliosides by Direct Infusion ESI-CID Tandem Mass Spectrometry of Native and Permethylated Molecular Species

Defining the Ceramide Composition of Bovine and Human Milk Gangliosides by Direct Infusion ESI-CID Tandem Mass Spectrometry of Native and Permethylated Molecular Species

Fucosylated glycoproteins and fucosylated glycolipids play opposing roles in cholera intoxication

Contact Info

Product

Resources

About