Proteomic analysis of glycosylphosphatidylinositol-anchored membrane proteins.
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Elortza F, Nuhse TS, Foster LJ, Stensballe A, Peck SC, Jensen ON
Proteomic analysis of glycosylphosphatidylinositol-anchored membrane proteins.
Mol Cell Proteomics. 2003 Dec;2(12):1261-70. Epub 2003 Sep 29.
- PubMed ID
- 14517339 [ View in PubMed]
- Abstract
Glycosylphosphatidylinositol-anchored proteins (GPI-APs) are a functionally and structurally diverse family of post-translationally modified membrane proteins found mostly in the outer leaflet of the plasma membrane in a variety of eukaryotic cells. Although the general role of GPI-APs remains unclear, they have attracted attention because they act as enzymes and receptors in cell adhesion, differentiation, and host-pathogen interactions. GPI-APs may represent potential diagnostic and therapeutic targets in humans and are interesting in plant biotechnology because of their key role in root development. We here present a general mass spectrometry-based proteomic "shave-and-conquer" strategy that specifically targets GPI-APs. Using a combination of biochemical methods, mass spectrometry, and computational sequence analysis we identified six GPI-APs in a Homo sapiens lipid raft-enriched fraction and 44 GPI-APs in an Arabidopsis thaliana membrane preparation, representing the largest experimental dataset of GPI-anchored proteins to date.