Subtype-specific trafficking of endothelin receptors

Yoichiro Abe, Kazuhisa Nakayama, Akihiro Yamanaka, Takeshi Sakurai, Katsutoshi Goto

Research output: Contribution to journalArticle

52 Citations (Scopus)

Abstract

We investigated the subcellular localization of two endothelin receptors (ET(A)R and ET(B)R). To visualize these receptors directly, the C terminus of each receptor was fused to the N terminus of enhanced green fluorescent protein (designated as ETR-EGFP). When transiently expressed in various mammalian cell lines, ETAR-EGFP was predominantly localized on the plasma membrane. By contrast, ET(B)R-EGFP was, independent of ligand stimulation, predominantly localized on the intracellular vesicular structures containing Lamp-1. Immunoblot analyses revealed that at steady state ET(B)R-EGFP was highly degraded, and its degradation was inhibited by bafilomycin A1. Antibody uptake experiments suggested that the ET(B)R-EGFP molecules were internalized from the plasma membrane. It is therefore likely that ET(B)R is first transported to the plasma membrane and then internalized, irrespective of ligand stimulation, to lysosomes where it undergoes proteolytic degradation. Exchanging the C-terminal cytoplasmic tails of the two ETRs revealed that the cytoplasmic tail is responsible for both the intracellular localization and the degradation of the receptors. Deletion of the extreme C- terminal 35 amino acids from both receptors allowed the receptor proteins to localize predominantly in the intracellular vesicles and to degrade. These observations indicate that the cytoplasmic tail of ET(A)R determines its plasma membrane localization. Stimulation with endothelin-1 increased the amount of intact ETR-EGFP fusion proteins without increasing their de novo synthesis, suggesting that binding of endothelin-1 stabilizes the ETRs.

Original languageEnglish
Pages (from-to)8664-8671
Number of pages8
JournalJournal of Biological Chemistry
Volume275
Issue number12
DOIs
Publication statusPublished - 2000 Mar 24

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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  • Cite this

    Abe, Y., Nakayama, K., Yamanaka, A., Sakurai, T., & Goto, K. (2000). Subtype-specific trafficking of endothelin receptors. Journal of Biological Chemistry, 275(12), 8664-8671. https://doi.org/10.1074/jbc.275.12.8664