S. Chatterjee et al., GPI anchoring leads to sphingolipid-dependent retention of endocytosed proteins in the recycling endosomal compartment, EMBO J, 20(7), 2001, pp. 1583-1592
Glycosylphosphatidylinositol (GPI) anchoring is important for the function
of several proteins in the context of their membrane trafficking pathways.
We have shown previously that endocytosed GPI-anchored proteins (GPI-APs) a
re recycled to the plasma membrane three times more slowly than other membr
ane components. Recently, we found that GPI-APs are delivered to endocytic
organelles, devoid of markers of the clathrin-mediated pathway, prior to th
eir delivery to a common recycling endosomal compartment (REC). Here we sho
w that the rate-limiting step in the recycling of GPI-APs is their slow exi
t from the REC; replacement of the GPI anchor with a transmembrane protein
sequence abolishes retention in this compartment. Depletion of endogenous s
phingolipid levels using sphingolipid synthesis inhibitors or in a sphingol
ipid-synthesis mutant cell line specifically enhances the rate of endocytic
recycling of GPI-APs to that of other membrane components. We have shown p
reviously that endocytic retention of GPI-APs is also relieved by cholester
ol depletion. These findings strongly suggest that functional retention of
GPI-APs in the REC occurs via their association with sphingolipid and chole
sterol-enriched sorting platforms or 'rafts'.