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Journal of Lipid Research, Vol 37, 2638-2649, Copyright © 1996 by Lipid Research, Inc.
MO Pentikainen, EM Lehtonen and PT Kovanen
In atherogenesis, low density lipoprotein (LDL, diameter 22 nm) accumulates
in the extracellular space of the arterial intima in the form of aggregates
of lipid droplets (droplet diameter up to 400 nm). Here we studied the
effects of various established in vitro LDL modifications on LDL
aggregation and fusion. LDL was subjected to vortexing, oxidation by copper
ions, proteolysis by alpha-chymotrypsin, lipolysis by sphingomyelinase, and
nonenzymatic glycosylation, and was induced to form adducts with
malondialdehyde or complexes with anti- apoB-100 antibodies. To assess the
amount of enlarged LDL-derived structures formed (due to aggregation or
fusion), we measured the turbidity of solutions containing modified LDL,
and quantified the proportion of modified LDL that 1) sedimented at
low-speed centrifugation (14,000 g), 2) floated at an increased rate at
high- speed centrifugation (rate zonal flotation at 285,000 gmax), 3) were
excluded in size-exclusion column chromatography (exclusion limit 40 MDa),
or 4) failed to enter into 0.5%. Fast Lane agarose gel during
electrophoresis. To detect whether particle fusion had contributed to the
formation of the enlarged LDL-derived structures, particle morphology was
examined using negative staining and thin-section transmission electron
microscopy. We found that 1) aggregation was induced by the formation of
LDL-antibody complexes, malondialdehyde treatment, and glycosylation of
LDL; 2) fusion of LDL was induced by proteolysis of LDL by
alpha-chymotrypsin; and 3) aggregation and fusion of LDL were induced by
vortexing, oxidation by copper ions, and lipolysis by sphingomyclinase of
LDL. The various modifications of LDL differed in their ability to induce
aggregation and fusion.
ARTICLES
Aggregation and fusion of modified low density lipoprotein
Wihuri Research Institute, Helsinki, Finland.
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