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MBC in Press, published online ahead of print February 21, 2007
Mol. Biol. Cell 10.1091/mbc.E06-10-0885

A more recent version of this article appeared on April 1, 2007
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Submitted on October 2, 2006
Revised on January 26, 2007
Accepted on February 1, 2007

Fatty Acid Remodeling of GPI-anchored Proteins Is Required for Their Raft Association

Yusuke Maeda,* Yuko Tashima,* Toshiaki Houjou,{dagger} Morihisa Fujita,{ddagger} Takehiko Yoko-o,{ddagger} Yoshifumi Jigami,{ddagger} Ryo Taguchi,{dagger} and Taroh Kinoshita*

*Research Institute for Microbial Diseases, Osaka University, Suita, Osaka 565-0871, Japan; {dagger}Graduate School of Medicine, University of Tokyo, Bunkyoku, Tokyo 113-0033, Japan; {ddagger}Research Center for Glycoscience, National Institute of Advanced Industrial Science and Technology, Tsukuba, Ibaraki 305-8566, Japan

Monitoring Editor: Reid Gilmore

Whereas most of the cellular phosphatidylinositol (PI) contain unsaturated fatty chains and are excluded from rafts, glycosylphosphatidylinositol-anchored proteins (GPI-APs) unusually contain two saturated fatty chains in their PI moiety and are typically found within lipid rafts. However, the origin of the saturated chains and whether they are essential for raft association are unclear. Here we report that GPI-APs, with two saturated fatty chains, are generated from those bearing an unsaturated chain by fatty acid remodeling that occurs most likely in the Golgi and requires PGAP2 and PGAP3. The surface GPI-APs isolated from the PGAP2&3-double-mutant CHO cells had unsaturated chains, such as oleic, arachidonic and docosatetraenoic acids in the sn-2 position; whereas those from wild-type CHO cells had exclusively stearic acid, a saturated chain, indicating that the sn-2 chain is exchanged to saturated one. We then assessed the association of GPI-APs with lipid rafts. Recovery of unremodeled GPI-APs from the double-mutant cells in the detergent-resistant membrane fraction was very low, indicating that GPI-APs become competent to be incorporated into lipid rafts by PGAP3- and PGAP2-mediated fatty acid remodeling. We also show that the remodeling requires the preceding PGAP1-mediated deacylation from inositol of GPI-APs in the ER.

Address correspondence to: Taroh Kinoshita (tkinoshi{at}biken.osaka-u.ac.jp)




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