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Glycobiology Advance Access published online on October 30, 2002
Glycobiology, doi:10.1093/glycob/cwg004
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Submitted on May 30, 2002
Revised on September 3, 2002
Accepted on September 3, 2002

© 2002 Oxford University Press

ORIGINAL ARTICLES

Structures of the glycosylphosphatidylinositol membrane anchors from Aspergillus fumigatus membrane proteins

Thierry Fontaine 1*, Thierry Magnin 1, Angela Melhert 2, Douglas Lamont 2, Jean-Paul Latgé 1, Michael A. J. Ferguson 2

1 Unité des Aspergillus, Institut Pasteur, 25 rue du Docteur Roux, 75724 Paris cedex 15, France
2 Division of Biological Chemistry & Molecular Microbiology, The School of Life Sciences, University of Dundee, Wellcome Trust Biocentre, Dow Street, Dundee DD1 5EH, UK

* To whom correspondence should be addressed. E-mail: tfontain{at}pasteur.fr.

Abstract

Glycosylphosphatidylinositol-anchored proteins have been identified in all eukaryotes. In the fungi, structural and biosynthetic studies of glycosylphosphatidylinositols have been restricted to the yeast Saccharomyces cerevesiae. In this study, four GPI-anchored proteins were purified from a membrane preparation of the human filamentous fungal pathogen Aspergillus fumigatus. Using new methodology applied to Western blot protein bands, the glycosylphosphatidylinositol structures were characterised by electrospray mass spectrometry, fluorescence labelling and high performance liquid chromatography and specific enzymatic digestions. The phosphatidylinositol moiety of the A.fumigatus glycosylphosphatidylinositol membrane anchors was shown to be an inositol-phosphoceramide containing mainly phytosphingosine and monohydroxylated C24:0 fatty acid. In constrast to yeast, only ceramide was found in the GPI-anchor structures of A. fumigatus, even for Gel1p, a homolog of Gas1p in S. cerevisiae that contains diacylglycerol. The A. fumigatus glycosylphosphatidylinositol glycan moiety is mainly a linear pentomannose structure linked to a glucosamine residue: Man{alpha}1-3Man{alpha}1-2Man{alpha}1-2Man{alpha}1-6Man{alpha}1-4GlcN.


Key words: Aspergillus / glycoprotein / glycosylphosphatidylinositol
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