Proteomic scale high-sensitivity analyses of GPI membrane anchors

Angela Mehlert, Michael A. J. Ferguson

    Research output: Contribution to journalArticle

    4 Citations (Scopus)

    Abstract

    Glycosylphosphatidylinositol (GPI) anchored proteins are ubiquitous in eukaryotic cells. Earlier analysis methods required large amounts of purified protein to elucidate the structure of the GPI. This paper describes methods for analyzing GPIs on a 'proteomic' scale. Partially purified proteins may be run on sodium dodecyl sulphate polyacrylamide gel electrophoresis and then blotted onto a polyvinylidene difluoride (PVDF) membrane. Following identification of the protein the piece of PVDF may be subjected to various chemical treatments, which are specific for GPI structures. The first method uses gas chromatography-mass spectrometry and it enables the presence of a GPI anchor to be confirmed. The second method depends on the cleavage of phosphate bonds and permits the carbohydrate structure to be elucidated by electrospray or matrix assisted laser desorption ionization-time of flight mass spectrometry. The final method described uses deamination of the glucosamine residue to release the lipid moiety for analysis by mass spectrometry.

    Original languageEnglish
    Pages (from-to)915-921
    Number of pages7
    JournalGlycoconjugate Journal
    Volume26
    Issue number8
    DOIs
    Publication statusPublished - Nov 2009

    Keywords

    • Glycosylphosphatidylinositol (GPI)
    • Mass spectrometry
    • Proteomic techniques
    • VARIANT SURFACE GLYCOPROTEIN
    • TRYPANOSOMA-BRUCEI
    • CRYPTOSPORIDIUM-PARVUM
    • ANTIGEN

    Cite this