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Journal of Lipid Research, Vol 36, 395-406, Copyright © 1995 by Lipid Research, Inc.
LL Swift
To investigate the assembly pathway for hepatic very low density
lipoproteins (VLDL), nascent lipoproteins were recovered from a purified,
intact rough endoplasmic reticulum (ER) fraction isolated from rat liver.
Two fractions were recovered by ultracentrifugation. Particles isolated at
d 1.006 g/ml were triglyceride-rich particles containing apolipoprotein
(apo)B-100 or apoB-48, and apoE with very small amounts of apoA-I. Compared
with VLDL recovered from the Golgi apparatus, the particles from the rough
ER had less triglyceride, but more cholesteryl ester and phospholipid. The
second class of particles isolated between d 1.006 and 1.210 g/ml were
phospholipid-rich and contained apoB-48, apoE, and apoA-I. ApoB-100 was a
minor component. Radioisotope incorporation studies utilizing [3H]leucine
revealed differential rates of labeling of the apoproteins in these two
lipoprotein fractions. ApoB-100 and apoE followed similar patterns in both
fractions with peak incorporation occurring within 15 min of isotope
injection. Incorporation of [3H]leucine into apoB-48 in the dense fraction
peaked within 15 min of isotope administration, but peak incorporation in
the d 1.006 g/ml fraction did not occur until approximately 30 min after
injection. We propose that the two lipoprotein fractions recovered from the
rough ER are intermediates in the assembly of VLDL by the liver. Comparison
of the composition of these two particles with that of Golgi VLDL supports
the sequential assembly of VLDL by the liver. Furthermore, we propose that
the initial steps in the assembly of apoB-100- and apoB-48-containing
lipoproteins are different with nascent apoB-100-containing particles being
formed through the cotranslational association of this apoprotein with
lipid while nascent apoB-48-containing VLDL are formed in the rough ER
through a two-step process.
ARTICLES
Assembly of very low density lipoproteins in rat liver: a study of nascent particles recovered from the rough endoplasmic reticulum
Department of Pathology, Vanderbilt University School of Medicine, Nashville, TN 37232-2561, USA.
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