SPHINGOLIPID ACTIVATOR PROTEIN-D (SAP-D) STIMULATES THE LYSOSOMAL DEGRADATION OF CERAMIDE IN-VIVO

Glycosphingolipids of cultured fibroblasts from patients with total sphingolipid activator proteins (SAPs) deficiency (Schnabel et al. J. Biol. Chem. 267:3312-3315, 1992) were labeled biosynthetically with [C-14]serine. After a chase period of 120h the patients' fibroblasts showed increased labeling of ceramide, glucosylceramide, lactosylceramide and ganglioside G(M3) in comparison to normal control cells. Addition of sap-D to the chase-media of the patients' fibroblasts led to the degradation of the accumulated ceramide down to nearly normal levels, whereas the levels of other labeled sphingolipids remained unaffected. In contrast, addition of sap-B to the chase-media of the patients' fibroblasts did not reduce the increased ceramide levels but resulted, as expected from in vitro experiments, in a specific decrease of levels of lactosylceramide and ganglioside G(M3) Therefore,we conclude that sap-D stimulates in vivo the lysosomal degradation of ceramide. (C) 1994 Academic Press, Inc.