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Requirement of conserved amino acid residues for ER‐export and recycling of platelet‐activating factor receptor
Author(s) -
Nakamura Motonao,
Hirota Nobuaki,
Yasuda Daisuke,
Hashidate Tomomi,
Yamaguchi Satoshi,
Nagamune Teruyuki,
Shimizu Takao
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.lb83
Subject(s) - internalization , endoplasmic reticulum , microbiology and biotechnology , alanine , endosome , receptor , amino acid , chemistry , biochemistry , biology
In the transmembrane domains (TMs) of G‐protein coupled receptors, several amino acid residues are conserved. Here, we demonstrate that a conserved proline, Pro 247 , in TM6 of platelet‐activating factor receptor (PAFR) is critical for endoplasmic reticulum (ER)‐export and recycling after agonist‐induced internalization. Alanine‐substituted mutants of the conserved residues of PAFRs, including P247A, were accumulated in the ER. Because a PAFR antagonist, Y‐24180, acted as a pharmacological chaperone to rescue ER‐retention, this retention is due to misfolding of PAFR. mc‐PAF, a PAFR agonist, did not increase surface expression of P247A, even if another ER‐retained mutant, D63A, was trafficked. Signaling and accumulation of the receptors in the early endosomes were observed in the mc‐PAF‐treated P247A‐expressing cells, suggesting that P247A was trafficked to the cell surface by mc‐PAF, thereafter internalized, but not recycled. Deficient recycling was confirmed with a sortase‐A‐mediated method for labeling cell surface proteins. These results demonstrate that the conserved proline in TM6 is crucial for the intracellular trafficking of PAFR. Supported in part by a Grants‐in‐aid from Ministry of Education, Culture, Sports, Science, and Technology of Japan