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Papers In Press, published online ahead of print November 19, 2001
Dept. Chemistry & Biochemistry, University of South Carolina, Columbia, SC 29208
Corresponding Author: wuthier{at}mail.chem.sc.edu
Matrix vesicles are lipid bilayer enclosed structures that initiate extracellular mineral formation and other functions related to bone development. 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 were taken for analysis of the lipids, extracted both before and after demineralization to remove deposited mineral. Phosphatidylserine and phosphatidylinositol both rapidly disappeared from extracts made before decalcification, indicative of possible 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.
J. Biol. Chem, 10.1074/jbc.M107899200
Submitted on August 16, 2001
Revised on November 15, 2001
Accepted on November 19, 2001
Changes in phospholipid extractability and composition accompany mineralization of chicken growth plate cartilage matrix vesicles
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