Application of electrospray mass spectrometry to the structural determination of glycosylphosphatidylinositol membrane anchors

The addition of glycosylphosphatidylinositol (GPI) anchors to proteins is an important posttranslational modification in eukaryotic cells. The complete structural elucidation of GPI anchors is a complex process that requires relatively large amounts of starting material. In this paper, we assess the...

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Bibliographic Details
Published in:Glycobiology (Oxford) 2010-05, Vol.20 (5), p.576-585
Main Authors: Nett, Isabelle R.E, Mehlert, Angela, Lamont, Douglas, Ferguson, Michael A.J
Format: Article
Language:English
Subjects:
glycosylphosphatidylinositol
Glycosylphosphatidylinositols - chemistry
GPI anchor
mass spectrometry
Molecular Structure
Original
Polysaccharides - chemistry
Spectrometry, Mass, Electrospray Ionization
Tandem Mass Spectrometry
Trypanosoma brucei
Trypanosoma brucei brucei - chemistry
variant surface glycoprotein
Variant Surface Glycoproteins, Trypanosoma - chemistry
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Summary:The addition of glycosylphosphatidylinositol (GPI) anchors to proteins is an important posttranslational modification in eukaryotic cells. The complete structural elucidation of GPI anchors is a complex process that requires relatively large amounts of starting material. In this paper, we assess the degree of structural information that can be obtained by applying electrospray mass spectrometry and tandem mass spectrometry to permethylated GPI glycans prepared from a well-characterized GPI-anchored glycoprotein, the variant surface glycoprotein from Trypanosoma brucei. All GPI glycans contain a non-N-acetylated glucosamine residue, and permethylation leads to the formation of a fixed positive charge on the glycans, in the form of a quaternary amine. The permethylated glycans were detected as [M +- Na]²⁺⁻ ions, and tandem mass spectrometry of these ions produced substantial, albeit incomplete, structural information on the branching patterns and linkage types for various GPI glycoforms of the variant surface glycoprotein.
ISSN:0959-6658
1460-2423
DOI:10.1093/glycob/cwq007