Complex sphingolipid profiling and identification of an inositol-phosphorylceramide synthase in Dictyostelium discoideum

Dictyostelium discoideum is a professional phagocyte frequently used to study cellular processes underlying the recognition, engulfment, and infection course of microbial pathogens. Sphingolipids are abundant components of the plasma membrane that bind cholesterol, control membrane properties, participate in signal transmission, and serve as adhesion molecules in recognition processes relevant to immunity and infection. By combining lipidomics with a bioinformatics-based cloning strategy, we show here that D. discoideum produces phosphoinositol-containing sphingolipids with predominantly phytoceramide backbones. Cell-free expression of candidate inositol-phosphorylceramide (IPC) synthases from D. discoideum enabled identification of an enzyme that selectively catalyzes the transfer of phosphoinositol from phosphatidylinositol onto ceramide. The IPC synthase, DdIPCS1, shares multiple sequence motifs with yeast IPC and human sphingomyelin synthases and localizes to the Golgi apparatus as well as the contractile vacuole of D. discoideum. These findings open up important opportunities for exploring a role of sphingolipids in phagocytosis and infection across major evolutionary boundaries. [Display omitted] •Dictyostelium discoideum produces phosphoinositol-containing sphingolipids•DdCSS2 shares structural features with inositol-phosphorylceramide (IPC) synthases•DdCSS2 shows IPC synthase activity•DdCSS2 is found in both, the Golgi apparatus and the contractile vacuole Biochemistry; Molecular biology; Cell biology; Bioinformatics; Lipidomics