Measurement of receptor endocytosis and recycling

Jane M. Knisely, Jiyeon Lee, Guojun Bu

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Receptor trafficking is essential to the delivery of nutrients and to the proper regulation of signaling pathways in mammalian cells. Numerous transmembrane receptors undergo clathrin-mediated endocytosis, followed by sorting in the early endosome. The low-density lipoprotein (LDL) receptor-related protein (LRP) is a multiligand endocytic receptor and a member of the LDL receptor family. At the cell surface, it binds to and continuously internalizes numerous ligands including lipoproteins, proteases, protease inhibitors, growth factors, and β-amyloid precursor protein via clathrin-mediated endocytosis. Its rapid endocytosis rate allows efficient clearance of extracellular and transmembrane ligands. Once internalized into the early or sorting endosome, LRP ligands are delivered to lysosomes to be degraded, whereas LRP is efficiently recycled to the plasma membrane. Herein, the authors describe quantitative methods to measure the endocytosis and recycling capacity of receptors, using LRP as a model receptor.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
Pages319-332
Number of pages14
Volume457
DOIs
StatePublished - 2008
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume457
ISSN (Print)10643745

Keywords

  • Endocytosis assay
  • Flow cytometry
  • Iodination
  • LRP
  • Quenching antibody
  • Recycling assay
  • TCA precipitation

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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  • Cite this

    Knisely, J. M., Lee, J., & Bu, G. (2008). Measurement of receptor endocytosis and recycling. In Methods in Molecular Biology (Vol. 457, pp. 319-332). (Methods in Molecular Biology; Vol. 457). https://doi.org/10.1007/978-1-59745-261-8-24