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Originally published In Press as doi:10.1074/jbc.M107899200 on November 19, 2001

J. Biol. Chem., Vol. 277, Issue 7, 5126-5133, February 15, 2002
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Changes in Phospholipid Extractability and Composition Accompany Mineralization of Chicken Growth Plate Cartilage Matrix Vesicles*

Licia N. Y. WuDagger , Brian R. GengeDagger , Min W. KangDagger , A. Larry Arsenault§, and Roy E. WuthierDagger

From the Dagger  Department of Chemistry and Biochemistry, University of South Carolina, Columbia, South Carolina 29208 and the § Department of Pathology, McMaster University, Hamilton, Ontario L8N 3Z5, Canada

Matrix vesicles are lipid bilayer-enclosed structures that initiate extracellular mineral formation. Little attention has been given to how newly formed mineral interacts with the lipid constituents and then emerges from the lumen. To explore whether specific lipids bind to the incipient mineral and if breakdown of the membrane is involved, we analyzed changes in lipid composition and extractability during vesicle-induced calcification. Isolated matrix vesicles were incubated in synthetic cartilage lymph to induce mineral formation. At various times, samples of the lipids were taken for analysis, extracted both before and after demineralization to remove deposited mineral. Phosphatidylserine and phosphatidylinositol both rapidly disappeared from extracts made before decalcification, indicating rapid degradation. However, extracts made after demineralization revealed that phosphatidylserine had become complexed with newly forming mineral. Concomitantly, its levels actually increased, apparently by base-exchange with phosphatidylethanolamine. Though partially complexed with the mineral, phosphatidylinositol was nevertheless rapidly broken down. Sphingomyelin and phosphatidylethanolamine also underwent rapid breakdown, but phosphatidylcholine was degraded more slowly, all accompanied by a buildup of free fatty acids. The data indicate that phosphatidylserine forms complexes that accompany mineral formation, while degradation of other membrane phospholipids apparently enables egress of crystalline mineral from the vesicle lumen.

* This work was supported by Grant AR18983 from the NIAMS, National Institutes of Health.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom correspondence should be addressed: Dept. of Chemistry and Biochemistry, Univ. of South Carolina, 329 Graduate Science Research Center, Columbia, SC 29208. Tel.: 803-777-6626; Fax: 803-777-9521; E-mail: wuthier@mail.chem.sc.edu.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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