Low density lipoprotein receptor-related protein mediates uptake of cholesteryl esters derived from apoprotein E-enriched lipoproteins.
Low density lipoprotein receptor-related protein (LRP) is a recently described cell-surface protein of 4544 amino acids that contains reiterated sequences found in the 839-amino acid receptor for low density lipoprotein (LDL). In the current studies, we purified LRP from rat liver, prepared polyclonal antibodies that recognize the extracellular domain, and demonstrated an immunoreactive protein of approximately 600 kDa in human fibroblasts. The function of this LRP was studied in mutant human fibroblasts that do not produce LDL receptors. The mutant cells were incubated with beta-migrating very low density lipoprotein (beta-VLDL) that was isolated from cholesterol-fed rabbits and artificially enriched with apoprotein (apo) E by incubation in vitro with human apo E produced in a bacterial expression system. The apo E-enriched beta-VLDL, but not unincubated beta-VLDL, stimulated incorporation of [14C]-oleate into cholesteryl [14C]oleate 20- to 40-fold in the mutant cells. This stimulation was blocked by chloroquine, suggesting that such stimulation resulted from receptor-mediated uptake and lysosomal hydrolysis of the cholesteryl esters in apo E-enriched beta-VLDL. Stimulation of cholesterol esterification was blocked by the antibody against LRP, but not by an antibody against the LDL receptor. Unlike the LDL receptor, the amount of LRP was not reduced when cells were incubated with oxygenated sterols. We conclude that LRP can mediate the cellular uptake and lysosomal hydrolysis of cholesteryl esters contained in lipoproteins that are enriched in apo E.
Department of Molecular Genetics, University of Texas Southwestern Medical Center, Dallas 75235.
R C
RC
Kowal
J
J
Herz
J L
JL
Goldstein
V
V
Esser
M S
MS
Brown
D000818:Animals; D001057:Apolipoproteins E / Q000378:metabolism*; D002478:Cells, Cultured; D002738:Chloroquine / Q000494:pharmacology; D002788:Cholesterol Esters / Q000378:metabolism*; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D008079:Lipoproteins, VLDL / Q000378:metabolism; D008099:Liver / Q000378:metabolism*; D026503:Low Density Lipoprotein Receptor-Related Protein-1; D008970:Molecular Weight; D009154:Mutation; D019301:Oleic Acid; D009829:Oleic Acids / Q000378:metabolism; D051381:Rats; D011971:Receptors, Immunologic / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D011973:Receptors, LDL / Q000096:biosynthesis / Q000378:metabolism*; D012867:Skin / Q000378:metabolism; D014280:Triglycerides / Q000096:biosynthesis
D001057:Apolipoproteins E; D002788:Cholesterol Esters; D008079:Lipoproteins, VLDL; D026503:Low Density Lipoprotein Receptor-Related Protein-1; D009829:Oleic Acids; D011971:Receptors, Immunologic; D011973:Receptors, LDL; D014280:Triglycerides; D019301:Oleic Acid; D002738:Chloroquine