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Title Proteomic scale high-sensitivity analyses of GPI membrane anchors
Edition Volume 26, Number 8
Call Number
ISBN/ISSN 0282-0080
Author(s) Mehlert,Angela
Ferguson,Michael A. J.
Subject(s)
Classification
Series Title Glycoconjugate Journal
GMD Electronic Journal
Language English
Publisher Springer Netherlands
Publishing Year 2009
Publishing Place Netherlands
Collation 7p
Abstract/Notes 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 ionizationtime
of flight mass spectrometry. The final method
described uses deamination of the glucosamine residue to
release the lipid moiety for analysis by mass spectrometry
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