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Journal of Lipid Research, Vol. 42, 1645-1654, October 2001
Copyright © 2001 by Lipid Research, Inc.

Original Article

N-linked glycosylation of macrophage-derived PAF-AH is a major determinant of enzyme association with plasma HDL

Correspondence to: Ewa Ninio, at INSERM U525, Faculté de Médecine Pitié-Salpêtrière, 91 Bd de l'Hôpital, 75634 Paris Cedex 13, France., ninio{at}chups.jussieu.fr (E-mail)

Human plasma PAF-AH (platelet-activating factor-acetylhydrolase) is a Ca²⁺-independent phospholipase A₂ of hematopoietic origin associated with LDL and HDL; it degrades PAF and oxidizes phospholipids. We show that human macrophages synthesize PAF-AH as a premedial Golgi precursor containing high mannose N-linked glycans. Secreted PAF-AH possesses a molecular mass of 55 kDa and contains mature N-linked glycans. Secreted PAF-AH activity (90 ± 4% of the total) bound to a wheat germ lectin column and could be eluted with N-acetylglucosamine, whereas digestion with N-acetylneuraminidase II completely abolished enzyme absorption. Tunicamycin significantly reduced cell-associated PAF-AH activity and inhibited enzyme secretion; but it did not alter the ratio of secreted to cell-associated enzyme (1.8 at 6 h and 3.1 at 24 h), suggesting that glycosylation is not essential for PAF-AH secretion. Digestion of cell-associated PAF-AH or secreted PAF-AH with peptide N-glycosidase F affected neither catalytic activity nor its resistance to proteolysis with trypsin or proteinase K; in addition, it did not affect PAF-AH association with LDL, but significantly increased its association with HDL.

We suggest that macrophage-derived PAF-AH contains heterogeneous asparagine-conjugated sugar chain(s) involving sialic acid, which hinders its association with HDL but does not influence the secretion, catalytic activity, or resistance of PAF-AH to proteases. — Tselepis, A. D., S-A. P. Karabina, D. Stengel, R. Piédagnel, M. J. Chapman, and E. Ninio. N-linked glycosylation of macrophage-derived PAF-AH is a major determinant of enzyme association with plasma HDL. J. Lipid Res. 2001. 42: 1645–1654.

Supplementary key words: atherogenesis, lipoproteins, macrophages

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