Abstract
Gangliosides, as plasma membrane-associated sialylated glycolipids, are antigenic structures and they serve as ligands for adhesion proteins of pathogens, for toxins of bacteria, and for endogenous proteins of the host. The detectability by carbohydrate-binding proteins of glycan antigens and ligands on glycolipids can be influenced by the differing lipid moieties. To investigate glycan sequences of gangliosides as recognition structures, we have underway a program of work to develop a “gangliome” microarray consisting of isolated natural gangliosides and neoglycolipids (NGLs) derived from glycans released from them, and each linked to the same lipid molecule for arraying and comparative microarray binding analyses. Here, in the first phase of our studies, we describe a strategy for high-sensitivity assignment of the tetrasaccharide backbones and application to identification of eight of monosialylated glycans released from bovine brain gangliosides. This approach is based on negative-ion electrospray mass spectrometry with collision-induced dissociation (ESI-CID-MS/MS) of the desialylated glycans. Using this strategy, we have the data on backbone regions of four minor components among the monosialo-ganglioside-derived glycans; these are of the ganglio-, lacto-, and neolacto-series.Graphical abstract
Highlights
Gangliosides are plasma membrane-associated and sialylated glycolipids
Sialylated glycans were released from extracted total bovine brain glycolipids by ozonolysis and alkaline hydrolysis
By negative-ion ESICID-mass spectrometry (MS)/MS [10, 17] analyses after desialylation, a wealth of sequence and linkage information is obtained at high sensitivity (1 pmol) on the backbone sequences of seven of the minor components among the monosialylated glycans released from bovine gangliosides
Summary
Gangliosides are plasma membrane-associated and sialylated glycolipids. The glycan moieties constitute. As the quantity obtainable can be too low for conventional NMR analysis, a high-sensitivity MS-based method is highly desirable In this presentation, we explore the negative-ion electrospray mass spectrometry with collisioninduced dissociation (ESI-CID-MS/MS) method, previously developed for both neutral [10, 11] and sialylated oligosaccharides [12], for assignment of the glycan backbone sequences of monosialylated gangliosides. We describe a new strategy for backbone assignment of monosialylated oligosaccharides released from bovine brain gangliosides This takes advantage of the wealth of sequence information obtainable from neutral oligosaccharides by negative-ion ESI-CID-MS/MS after chemical desialylation. Using this strategy, we have identified the major and some of the very minor components of bovine brain gangliosides. Solvent (CH3CN/2 mM NH4HCO3 1:1) was delivered by a syringe pump at a flow rate of 10 μl/min
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callMore From: Journal of the American Society for Mass Spectrometry
Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
