Formation of phospholipid-rich HDL: a model for square-packing lipoprotein particles found in interstitial fluid and in abetalipoproteinemic plasma

Abstract

The major bovine HDL subfraction, fraction I-HDL, was incubated with increasing amounts of dimyristoylphosphatidylcholine (DMPC). HDL size, as determined by gradient gel electrophoresis and electron microscopy, increased with increasing HDL-phospholipid to DMPC mole ratios. Control fraction I-HDL were spherical, hexagonally-packing particles with a peak on gradient gel electrophoresis at 12.3 ± 0.1 nm; at a ratio of 1:0.5, larger, mainly spherical particles with a peak at 12.9 ± 0.08 nm were formed. At a ratio of 1:1, occasional square-shaped particles were seen by electron microscopy; by gradient gel analysis, the mean diameter of the HDL-product increased to 13.7 ± 0.1 nm. At the 1:2 ratio, extensive domains of squarepacking particles were noted; the major size peak of this product was 14.6 ± 0.08 nm. In all incubations with DMPC, a small 9.4 ± 0.08 nm product was formed; it was most pronounced at the 1:2 ratio. The large, less dense particles generated by incubation contained apolipoprotein A-I and small molecular weight proteins. The 9.4 nm product contained only apolipoprotein A-I. The less dense product formed during incubation at the 1:2 ratio had a decreased protein-to-lipid ratio relative to control HDL and a 2-fold increase in percent phospholipid. At a 1:2 ratio, incorporation of DMPC into fraction I-HDL results in the loss of one molecule of apolipoprotein A-I; the resultant particle is a stable phospholipid-rich and protein-poor HDL which has a square-packing geometry. These phospholipid-laden HDL are morphologically similar to lipoproteins isolated from interstitial fluid or from plasma of abetalipoproteinemic patients. Our data suggest that the unusual morphological properties of the latter biologically formed particles may be due to increases in the polar lipid contents, and concomitant decreases in surface protein.