Isolation of a glycosyl-phosphatidylinositol (GPI) from rat brain

Abstract

Brain lipids were labelled with [1-14C]-isethionyl acetimidate and purified by sequential thin layer chromatography. Four labelled peaks were obtained, the first ones migrating with the same Rf as glycosyl-phosphatidylinositol (GPI). Further proof of the isolation of GPI was obtained by the observations that 44.8% of the radioactivity associated with the lipid in peak I was converted to the water phase by the effect of a PI-specific phospholipase C, and that the soluble material so obtained produced a dose-dependent inhibition of cAMP-dependent protein kinase activity. These findings indicate a biological equivalence between GPI and its polar head group from rat brain and those described in other cell types, and are consistent with the proposed role of these molecules in cellular signalling.