Characterization in vitro of interaction of human apolipoprotein E-free high density lipoprotein with human hepatocytes

D. Schouten, M. F. Kleinherenbrink-Stins, A. Brouwer, D. L. Knook, J. A.A.M. Kamps, J. Kuiper, T. J.C. Van Berkel

Research output: Contribution to journalArticlepeer-review

Abstract

Characterization of the interaction of iodinated apolipoprotein (apo) E-free high density lipoprotein (HDL) with cultured human hepatocytes provides evidence for a saturable, Ca2+-independent, high affinity binding site with an apparent k(m) value of 20 μg/ml of apolipoprotein. Nitrated HDL and low density lipoprotein (LDL) did not compete for the binding of HDL, in contrast to very low density lipoprotein (VLDL). It is suggested that VLDL competition is exerted by the presence of apo Cs. Degradation of HDL was relatively low and in some cases not detectable. In cases where degradation was found, inhibitors of the lysosomal pathway of protein degradation had no effect, while LDL degradation was inhibited more than 80%. In the presence of 10 μM of monensin, the cell-association of HDL was unaffected, but the degradation was inhibited by 30%. Under similar conditions, LDL association was inhibited by 40% and LDL degradation by 90%. Incubation of human hepatocytes with fluorescently labeled HDL (Dil-HDL) revealed (in contrast to Dil-LDL) mainly strong membrane-bound fluorescence and hardly any labeling of small intracellular vesicles. It is concluded that human hepatocytes possess a specific high affinity site for human HDL with recognition properties similar to those described earlier on rat hepatocytes. No evidence that the binding of HDL is actively coupled to uptake and lysosomal degradation could be obtained, indicating that binding of LDL and HDL to human hepatocytes is coupled differently to intracellular pathways.

Original languageEnglish (US)
Pages (from-to)1127-1135
Number of pages9
JournalArteriosclerosis
Volume10
Issue number6
DOIs
StatePublished - 1990
Externally publishedYes

Keywords

  • high density lipoproteins
  • human hepatocytes
  • human parenchymal cells
  • lipoproteins

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine

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