Integrated mass spectrometric strategy for characterizing the glycans from glycosphingolipids and glycoproteins: direct identification of sialyl Le(x) in mice.
The current interest in applying systems biology approaches to studying an organism's form or function promises to reveal further insights into the role of glycosylation in cells and whole organisms. This has prompted the development of a rapid, sensitive method of profiling the glycan component of both glycosphingolipids and glycoproteins from a single sample. Here we report a new mass spectrometric screening strategy for characterizing glycosphingolipid-derived oligosaccharides, which can be integrated into an existing highly sensitive glycoprotein glycomics strategy. Using ceramide glycanase to release the glycans from glycosphingolipids, this method provides a reliable profile of the glycosphingolipid-derived glycans present in a sample and has revealed new glycan structures. Glycoproteins are also efficiently recovered using this method, allowing the subsequent analysis of glycoprotein-derived glycans by mass spectrometry. The high sensitivity of this glycomic screening method allowed us to directly characterize the sialyl Le(x) epitope from mouse brain for the first time, where it was observed on an O-mannose structure. Thus, we present a mass spectrometric method that allows glycomic screening of N- and O-glycans as well as glycosphingolipid-derived glycans from a single tissue.
Parry
Simon
S
Division of Molecular Biosciences, Imperial College London, London SW7 2AZ, UK.
Ledger
Victoria
V
Tissot
Bérangère
B
Haslam
Stuart M
SM
Scott
James
J
Morris
Howard R
HR
Dell
Anne
A
D000818:Animals; D001923:Brain Chemistry; D002240:Carbohydrate Sequence; D006023:Glycoproteins / Q000737:chemistry*; D006026:Glycoside Hydrolases / Q000494:pharmacology; D006028:Glycosphingolipids / Q000737:chemistry*; D006031:Glycosylation; D006168:Guinea Pigs; D008297:Male; D013058:Mass Spectrometry / Q000379:methods*; D051379:Mice; D008810:Mice, Inbred C57BL; D008815:Mice, Inbred Strains; D009844:Oligosaccharides / Q000032:analysis* / Q000737:chemistry; D011134:Polysaccharides / Q000032:analysis* / Q000737:chemistry; D051381:Rats; D017207:Rats, Sprague-Dawley; D012680:Sensitivity and Specificity; D000080506:Sialyl Lewis X Antigen; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013997:Time Factors; D014018:Tissue Distribution
D006023:Glycoproteins; D006028:Glycosphingolipids; D009844:Oligosaccharides; D011134:Polysaccharides; D000080506:Sialyl Lewis X Antigen; D006026:Glycoside Hydrolases; C051111:ceramide glycanase