High-Affinity Uptake and Degradation of Apolipoprotein E Free High-Density Lipoprotein and Low-Density Lipoprotein in Cultured Porcine Hepatocytes

Paul S. Bachorik, Frank A. Franklin, Donna G. Virgil, Peter O. Kwiterovich

Research output: Contribution to journalArticlepeer-review

Abstract

Isolated pig liver membranes contain a specific “lipoprotein binding site” that recognizes low-density lipoprotein (LDL) and apolipoprotein E (apoE) free high-density lipoprotein (HDL) [Bachorik, P. S., Kwiterovich, P. O., & Cooke, J. (1978) Biochemistry 17, 5287–5299]. We report here that a similar site exists in cultured porcine hepatocytes and that it mediates the uptake and degradation of apoE-free HDL. The binding of 125I-labeled HDL and 125I-labeled LDL (125I-HDL and 125I-LDL, respectively) at 4 °C and the uptake and degradation of the lipoproteins at 37 °C were time dependent and saturable and were not inhibited by unrelated proteins. Chloroquine (6×10−5 M) inhibited the degradation of 125I-HDL by 76% and of 125I-LDL by >99%; leupeptin inhibited the degradation of both lipoproteins by about 25%. I25I-HDL binding (4 °C), uptake, and degradation (37 °C) were inhibited by LDL, methyl-LDL, and methyl-HDL about as well as by unlabeled HDL but were unaltered in Pronase-treated cells or in cells that were cultured for 24 h in either lipoprotein-free medium or medium containing HDL or LDL (200 µg/mL). In contrast, these conditions affected the uptake and degradation of 125I-LDL disproportionately. HDL and methyl-LDL inhibited 125I-LDL uptake by 50% or more but had little effect on degradation. 125I-LDL binding was reduced by 12% and degradation by 57% in Pronase-treated cells. Preincubation of the cells with LDL (200 µg/mL) reduced uptake by 35% and degradation by 68%. Similar preincubation with HDL (200 µg/mL) increased 125I-LDL degradation by 60% but did not affect 125I-LDL uptake. The findings indicated the presence in porcine hepatocytes of at least two distinct sites for lipoproteins. One site resembled the LDL receptor and mediated 125I-LDL degradation. A second, Pronase-insensitive site recognized both HDL and LDL. This site mediated almost all of the degradation of 125I-HDL but little if any degradation of 125I-LDL.

Original languageEnglish (US)
Pages (from-to)5675-5684
Number of pages10
JournalBiochemistry
Volume21
Issue number22
DOIs
StatePublished - Jan 1 1982

ASJC Scopus subject areas

  • Biochemistry

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