Cholesterol-regulated translocation of NPC1L1 to the cell surface facilitates free cholesterol uptake

Liqing Yu, Shantaram Bharadwaj, J. Mark Brown, Yinyan Ma, Wei Du, Matthew A. Davis, Peter Michaely, Pingsheng Liu, Mark C. Willingham, Lawrence L. Rudel

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Abstract

Although NPC1L1 is required for intestinal cholesterol absorption, data demonstrating mechanisms by which this protein facilitates the process are few. In this study, a hepatoma cell line stably expressing human NPC1L1 was established, and cholesterol uptake was studied. A relationship between NPC1L1 intracellular trafficking and cholesterol uptake was apparent. At steady state, NPC1L1 proteins localized predominantly to the transferrin-positive endocytic recycling compartment, where free cholesterol also accumulated as revealed by filipin staining. Interestingly, acute cholesterol depletion induced with methyl-β-cyclodextrin stimulated relocation of NPC1L1 to the plasma membrane, preferentially to a newly formed "apical-like" subdomain. This translocation was associated with a remarkable increase in cellular cholesterol uptake, which in turn was dose-dependently inhibited by ezetimibe, a novel cholesterol absorption inhibitor that specifically binds to NPC1L1. These findings define a cholesterol-regulated endocytic recycling of NPC1L1 as a novel mechanism regulating cellular cholesterol uptake.

Original languageEnglish (US)
Pages (from-to)6616-6624
Number of pages9
JournalJournal of Biological Chemistry
Volume281
Issue number10
DOIs
StatePublished - Mar 10 2006

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ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Yu, L., Bharadwaj, S., Brown, J. M., Ma, Y., Du, W., Davis, M. A., Michaely, P., Liu, P., Willingham, M. C., & Rudel, L. L. (2006). Cholesterol-regulated translocation of NPC1L1 to the cell surface facilitates free cholesterol uptake. Journal of Biological Chemistry, 281(10), 6616-6624. https://doi.org/10.1074/jbc.M511123200