Structures of glycosylphosphatidylinositol membrane anchors from Saccharomyces cerevisiae.

Metabolic labeling studies suggest that Saccharomyces cerevisiae contains many glycoproteins that are anchored in the lipid bilayer by glycosylphosphatidylinositol membrane anchors. Membrane anchors were purified from a crude yeast membrane protein fraction and analyzed by two-dimensional 1H-1H NMR, fast atom bombardment-mass spectrometry, compositional and methylation linkage analyses, as well as chemical and enzymatic modifications. The yeast glycosylphosphatidylinositol anchors consist of the following structures: ethanolamine-PO4-6(R-2)Man alpha 1-2Man alpha 1-6Man alpha 1-4Glc-NH2 alpha 1-6myo-inositol-1-PO4-lipid, where R is mainly Man alpha 1- (80%) with some Man alpha 1-2Man alpha 1- (15%) and Man alpha 1-3Man alpha 1- (5%). The core region of the yeast anchors (ethanolamine-PO4-6Man alpha 1-2Man alpha 1-6Man alpha 1-4GlcNH2 alpha 1-6myo-inositol-1-PO4) is identical to the conserved core region found in glycosylphosphatidylinositol anchors from protozoa and mammals. The lipid moieties of the total yeast glycosylphosphatidylinositol anchors are mainly ceramides, consisting mostly of C18:0 phytosphingosine and C26:0 fatty acid. However, the lipid moiety of the glycosylphosphatidylinositol anchor of the purified ggp125 protein is a lyso- or diacylglycerol, containing C26:0 fatty acids. This suggests that yeast adds different lipid components to the glycosylphosphatidylinositol anchors of different proteins.