Intracellular trafficking of recycling apolipoprotein E in Chinese hamster ovary cells

Nicole A. Braun, Peter J. Mohler, Karl H. Weisgraber, Alyssa H. Hasty, MacRae F. Linton, Patricia G. Yancey, Ru Su Yan, Sergio Fazio, Larry L. Swift

Research output: Contribution to journalArticlepeer-review

15 Scopus citations

Abstract

We have investigated apolipoprotein E (apoE) recycling in Chinese hamster ovary (CHO) cells, a peripheral cell that does not produce lipoproteins or express apoE. Using a pulse-chase protocol in which cells were pulsed with 125I-apoE-VLDL and chased for different periods, ∼30% of the apoE internalized during the pulse was resecreted within a 4 h chase in a relatively lipid-free state. The addition of lysosomotropic agents or brefeldin A had no effect on apoE recycling. Unlike previous results with hepatocytes and macrophages, neither apoA-I nor upregulation of ABCA1 stimulated apoE recycling. However, cyclodextrin, which extracts cholesterol from plasma membrane lipid rafts, increased recycling. Confocal studies revealed that apoE, internalized during a 1 h pulse, colocalizes with early endosomal antigen-1, Rab5, Rab11a, and lysobisphosphatidic acid but not with lysosomal-associated membrane protein-1. Colocalization of apoE and Rab11a persisted even after cells had been chased for 1 h, suggesting a pool of apoE within the endosomal recycling compartment (ERC). Our data suggest that apoE recycling in CHO cells is linked to cellular cholesterol removal via the ERC and phospholipid-containing acceptors in a pathway alternative to the ABCA1-apoA-I axis.

Original languageEnglish (US)
Pages (from-to)1176-1186
Number of pages11
JournalJournal of lipid research
Volume47
Issue number6
DOIs
StatePublished - Jun 2006
Externally publishedYes

Keywords

  • ATP binding cassette transporter A1
  • Cholesterol efflux
  • Early endosomes
  • Rab11a
  • Rab5
  • Recycling endosomes
  • β-cyclodextrin

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology
  • Cell Biology

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