期刊
JOURNAL OF MASS SPECTROMETRY
卷 45, 期 5, 页码 504-519出版社
WILEY
DOI: 10.1002/jms.1734
关键词
Glycosphingolipid; sphingolipid; ganglioside; glycosylinositol phosphorylceramide; mass spectrometry; MALDI-TOF-MS; ESI-MS; linear ion trap; microarray; glycomics; sphingolipidomics
资金
- NIH [R21 RR20355]
- Copenhagen Center for Glycomics at the University of Copenhagen
- Danish Medical Research Council for Technology and Innovation
- Stjerne Program of Excellence
- University of New Hampshire Center for Structural Biology (NIH/NCRR) [P20 8R16459]
A method for generation of novel fluorocarbon derivatives of glycosphingolipids (GSLs) with high affinity for fluorocarbon phases has been developed, and their potential applications to mass spectrometry (MS)-based methodologies for glycosphingolipidomics have been investigated. Sphingolipid ceramide N-deacylase (SCDase) is used to remove the fatty acid from the ceramide moiety, after which a fluorocarbon-rich substituent (F-Tag) is incorporated at the free amine of the sphingoid. In initial trials, a neutral GSL, globotriaosylceramide (Gb(3)Cer), three purified bovine brain gangliosides, and four fungal glycosylinositol phosphorylceramides (GIPCs) were de-N-acylated, derivatized by prototype F-Tags, and recovered by solid phase extraction on fluorocarbon-derivatized silica (F-SPE). The efficacy of SCDase treatment of GIPCs was here demonstrated for the first time. Compatibility with subsequent per-N,O-methylation was established for the F-tagged Gb(3) Cer and purified gangliosides, and extensive mass spectra (MS1 and MS2) consistent with all of the expected products were acquired. The potential use of F-tagged derivatives for a comprehensive MS based profiling application was then demonstrated on a crude ganglioside mixture extracted from bovine brain. Finally, a simple trial in microarray format demonstrated fixation of F-tagged G(M1) ganglioside to a fluorous glass surface, with the glycan intact and available for interaction with a fluorescent derivative of cholera toxin B chain. The methods described thus provide a new avenue for rapid GSL recovery or cleanup, potentially compatible with a variety of platforms for mass spectrometric profiling and structure analysis, as well as parallel analysis of functional interactions. Copyright (C) 2010 John Wiley & Sons, Ltd.
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