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Journal of Lipid Research, Vol. 48, 1599-1606, July 2007
Copyright © 2007 by American Society for Biochemistry and Molecular Biology

Changes in molecular species profiles of glycosylphosphatidylinositol anchor precursors in early stages of biosynthesis

Toshiaki Houjou^*, Jun Hayakawa^1,, Reika Watanabe^2,, Yuko Tashima, Yusuke Maeda, Taroh Kinoshita and Ryo Taguchi^3,*,

^* Graduate School of Medicine, University of Tokyo, Bunkyo-ku, Tokyo 113-0033, Japan
Nagoya City University, Nagoya-shi, Aichi 467-8603, Japan
Research Institute for Microbial Diseases, Osaka University, Suita-shi, Osaka 565-0871, Japan

Published, JLR Papers in Press, April 20, 2007.

¹ Present address of J. Hayakawa: Shionogi Co., Ltd., 3-1-8 Docyuumachi, Cyuoh-Ku, Osaka Japan.

² Present address of R. Watanabe: Department of Biochemistry, Sciences II, 30 Quai E. Ansermet, CH-1211 Geneva 4, Switzerland.

³ To whom correspondence should be addressed. e-mail: rytagu{at}m.u-tokyo.ac.jp

Glycosylphosphatidylinositol (GPI) anchor is a major lipidation in posttranslational modification. GPI anchor precursors are biosynthesized from endogenous phosphatidylinositols (PIs) and attached to proteins in the endoplasmic reticulum. Endogenous PIs are characterized by domination of diacyl species and the presence of polyunsaturated fatty acyl chain, such as 18:0-20:4, at the sn-2 position. In contrast, the features of mammalian glycosylphosphatidylinositol-anchored proteins (GPI-APs) are domination of alkyl/acyl PI species and the presence of saturated fatty acyl chains at the sn-2 position, the latter being consistent with association with lipid rafts. Recent studies showed that saturated fatty acyl chain at sn-2 is introduced by fatty acid remodeling that occurs in GPI-APs. To gain insight into the former feature, we analyzed the molecular species of several different GPI precursors derived from various mammalian mutant cell lines. Here, we show that the PI species profile greatly changed in the precursor glucosamine (GlcN)-acyl-PI and became very similar to that of GPI-APs before fatty acid remodeling. They had alkyl (or alkenyl)/acyl types with unsaturated acyl chain as the major PI species. Therefore, a specific feature of the PI moieties of mature GPI-APs, domination of alkyl (or alkenyl)/acyl type species over diacyl types, is established at the stage of GlcN-acyl-PI.

Supplementary key words lipid molecular species • fatty acyl chain remodeling • mass spectrometry • liquid chromatography-electrospray ionization-mass spectrometry • phospholipid localization

Abbreviations: alk/acyl, alkyl/acyl or alkenyl/acyl; f.a., fatty acyl residue; GlcN, glucosamine; GlcNAc, N-acetyl glucosamine; GPI, glycosylphosphatidylinositol; GPI-AP, glycosylphosphatidylinositol-anchored protein; Ins-P, inositol phosphate; LC-ESI-MS, liquid chromatography-electrospray ionization-mass spectrometry; PGAP, Post-GPI Attachment to Proteins; PI, phosphatidylinositol

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