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Lip1p: a novel subunit of acyl‐CoA ceramide synthase
Author(s) -
Vallée Béatrice,
Riezman Howard
Publication year - 2005
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.1038/sj.emboj.7600562
Subject(s) - ceramide , ceramide synthase , sphingolipid , biology , biochemistry , atp synthase , protein subunit , enzyme , lipid signaling , biosynthesis , gene , apoptosis
Ceramide plays a crucial role as a basic building block of sphingolipids, but also as a signalling molecule mediating the fate of the cell. Although Lac1p and Lag1p have been shown recently to be involved in acyl‐CoA‐dependent ceramide synthesis, ceramide synthase is still poorly characterized. In this study, we expressed tagged versions of Lac1p and Lag1p and purified them to near homogeneity. They copurified with ceramide synthase activity, giving unequivocal evidence that they are subunits of the enzyme. In purified form, the acyl‐CoA dependence, fatty acyl‐CoA chain length specificity, and Fumonisin B1/Australifungin sensitivity of the ceramide synthase were the same as in cells, showing that these are properties of the enzyme and do not depend upon the membrane environment or other factors. SDS–PAGE analysis of purified ceramide synthase revealed the presence of a novel subunit of the enzyme, Lip1p. Lip1p is a single‐span ER membrane protein that is required for ceramide synthesis in vivo and in vitro . The Lip1p regions required for ceramide synthesis are localized within the ER membrane or lumen.