Title : Mammalian Lass6 and its related family members regulate synthesis of specific ceramides
Abstract :
- The Lass (longevity-assurance homologue) family members, which are highly conserved among eukaryotes, function in ceramide synthesis
- In the mouse, there are at least five Lass family members, Lass1 , Lass2 , Lass4 , Lass5 and the hitherto uncharacterized Lass6
- To investigate specific roles for each Lass member in ceramide synthesis, we cloned these five mouse proteins
- Overproduction of any Lass protein in cultured cells resulted in an increase in cellular ceramide, but the ceramide species produced varied
- Overproduction of Lass1 increased C18:0-ceramide levels preferentially, and overproduction of Lass2 and Lass4 increased levels of longer ceramides such as C22:0- and C24:0-ceramides
- Lass5 and Lass6 produced shorter ceramide species (C14:0- and C16:0-ceramides); however, their substrate preferences towards saturated/unsaturated fatty acyl-CoA differed
- In addition to differences in substrate preferences, we also demonstrated by Northern blotting that Lass family members are differentially expressed among tissues
- Additionally, we found that Lass proteins differ with regard to glycosylation
- Of the five members, only Lass2 , Lass5 and Lass6 were N-glycosylated, each at their N-terminal Asn residue
- The occurrence of N-glycosylation of some Lass proteins provides topological insight, indicating that the N-termini of Lass family members probably face the luminal side of the endoplasmic reticulum membrane
- Furthermore, based on a proteinase K digestion assay, we demonstrated that the C-terminus of Lass6 faces the cytosolic side of the membrane
- From these data we propose topology for the conserved Lag1 motif in Lass family members, namely that the N-terminal region faces the luminal side and the C-terminal region the cytosolic side of the endoplasmic reticulum membrane
Output (sent_index, trigger,
protein,
sugar,
site):
- 10. N-glycosylation, , proteins, -, -
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*Output_Site_Fusion* (sent_index,
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