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Journal of Lipid Research, Vol 31, 103-112, Copyright © 1990 by Lipid Research, Inc.
DL Brasaemle and AD Attie
The kinetics of low density lipoprotein (LDL) cholesterol transport to the
plasma membrane of Chinese hamster ovary (CHO) cells was studied. LDL was
reconstituted with [3H]cholesteryl linoleate and added to CHO cells in a
pulse-chase experiment. The internalization and lysosomal cleavage of
reconstituted LDL (rLDL) [3H]cholesteryl linoleate to free [3H]cholesterol
occurred with a half-time of 37 min after a 30-min lag. The rate of
transport of released [3H]cholesterol to the plasma membrane was measured
by brief (20-30 sec) cholesterol oxidase treatment of intact, adherent
cells: the half-time of transport was 42 min. The similarity in the rate of
free cholesterol release from rLDL and transport of this cholesterol to the
plasma membrane suggests very rapid transport of rLDL cholesterol from the
lysosome to the plasma membrane. Cells were shown to be intact throughout
the cholesterol oxidase treatment by the absence of cell-derived lactate
dehydrogenase (LDH) activity or K+ in the assay buffer.
ARTICLES
Rapid intracellular transport of LDL-derived cholesterol to the plasma membrane in cultured fibroblasts
Department of Biochemistry, University of Wisconsin, Madison 53706.
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