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Journal of Lipid Research, Vol. 42, 442-451, March 2001
Copyright © 2001 by Lipid Research, Inc.

Original Article

GPI-specific phospholipase D associates with an apoA-I- and apoA-IV-containing complex

Correspondence to: Mark A. Deeg, at Endocrinology (111E), Roudebush VAMC, 1481 W. 10th Street, Indianapolis, IN 46202., mdeeg{at}iupui.edu (E-mail)

Glycosylphosphatidylinositol-specific phospholipase D (GPI-PLD) is abundant in serum and associates with high density lipoproteins (HDL). We have characterized the distribution of GPI-PLD among lipoproteins in human plasma. Apolipoprotein (apo)-specific lipoproteins containing apoB (Lp[B]), apoA-I and A-II (Lp[A-I, A-II]), or apoA-I only (Lp[A-I]) were isolated using dextran sulfate and immunoaffinity chromatography. In six human plasma samples with HDL cholesterol ranging from 39 to 129 mg/dl, 79 ± 14% (mean ± SD) of the total plasma GPI-PLD activity was associated with Lp[A-I], 9 ± 12% with Lp[A-I, A-II], and 1 ± 1% with Lp[B]; and 11 ± 10% was present in plasma devoid of these lipoproteins. Further characterization of the GPI-PLD-containing lipoproteins by gel-filtration chromatography and nondenaturing polyacrylamide and agarose gel electrophoresis revealed that these apoA-I-containing particles/complexes were small (8 nm) and migrated with pre-ß particles on agarose electrophoresis.

Immunoprecipitation of GPI-PLD with a monoclonal antibody to GPI-PLD co-precipitated apoA-I and apoA-IV but little or no apoA-II, apoC-II, apoC-III, apoD, or apoE. In vitro, apoA-I but not apoA-IV or bovine serum albumin interacted directly with GPI-PLD, but did not stimulate GPI-PLD-mediated cleavage of a cell surface GPI-anchored protein. Thus, the majority of plasma GPI-PLD appears to be specifically associated with a small, discrete, and minor fraction of lipoproteins containing apoA-I and apoA-IV. — Deeg, M. A., E. L. Bierman, and M. C. Cheung. GPI-specific phospholipase D associates with an apoA-I- and apoA-IV-containing complex. J. Lipid Res. 2001. 42: 442;–451.

Supplementary key words: lipoproteins, glycosylphosphatidylinositol, lipids, cholesterol, phospholipase D

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