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Journal of Lipid Research, Vol 32, 529-543, Copyright © 1991 by Lipid Research, Inc.
Isolation and properties of nascent lipoproteins from highly purified rat hepatocytic Golgi fractions
RL Hamilton, A Moorehouse and RJ Havel
Cardiovascular Research Institute, University of California, San Francisco 94143-0130.
Two procedures were used to isolate hepatocytic Golgi fractions from rat
liver. One procedure yields a light Golgi fraction (GF1 + 2) and the other
"intact" stacks of cisternae. Triglyceride fatty acids in nascent very low
density lipoproteins (VLDL) were labeled by injection of [3H]palmitate
intravenously, and radiolabeled lipoproteins were injected as markers of
potentially contaminating endosomes. GF1 + 2 fractions were enriched
manyfold in the endosomal markers, indicative of substantial endosomal
contamination, whereas intact Golgi fractions from the same livers were
about 7% as contaminated. By electron microscopy, GF1 + 2 fractions
contained mainly multivesicular bodies (MVBs), together with some
Golgi-derived secretory vesicles. The small endosomal contamination of
intact Golgi fractions was further reduced by a simple modification of the
procedure, which removed most entrained endosomes. The surface constituents
of Golgi VLDL (d less than 1.010 g/ml) released from these highly purified
intact Golgi fractions differed from those of plasma VLDL. Golgi VLDL
contained fivefold less unesterified cholesterol than plasma VLDL, but
twofold more phospholipids. Golgi VLDL and plasma VLDL contained similar
amounts of cholesteryl esters and triglycerides. The protein content of
Golgi VLDL was substantially lower than that of plasma VLDL. ApoB-100 and
apoB-48 were similarly represented, but nascent VLDL contained less of the
C apolipoproteins. ApoA-I was present mainly as the proprotein in Golgi
VLDL, but was virtually lacking in plasma VLDL. ApoE comprised about 22% of
the protein mass of Golgi VLDL as well as plasma VLDL; the distribution of
apoE isoforms was also similar. Apolipoproteins E and pro A-I released from
ruptured Golgi cisternae were largely bound to the Golgi VLDL or were
associated with Golgi membranes. Particles resembling low density
lipoproteins (LDL) and high density lipoproteins (HDL) were not seen by
electron microscopy in contents of intact Golgi fractions. These
observations indicate that nascent Golgi VLDL are the primary particulate
precursors of rat plasma lipoproteins of hepatocytic origin, and suggest
that particles with the density of plasma HDL and LDL do not exist within
the secretory pathway of normal hepatocytes. Thus, the results of this
research on the properties of nascent plasma lipoprotein precursors
contained within uncontaminated hepatocytic Golgi fractions differ
substantially from previous published work.

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Copyright © 1991 by the American Society for Biochemistry and Molecular Biology.
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