Chemerin C9 peptide induces receptor internalization through a clathrin-independent pathway
Zhou, JX; Liao, D; Zhang, S; Cheng, N; He, HQ; Ye, RD
HERO ID
2902771
Reference Type
Journal Article
Year
2014
Language
English
PMID
| HERO ID | 2902771 |
|---|---|
| In Press | No |
| Year | 2014 |
| Title | Chemerin C9 peptide induces receptor internalization through a clathrin-independent pathway |
| Authors | Zhou, JX; Liao, D; Zhang, S; Cheng, N; He, HQ; Ye, RD |
| Journal | Acta Pharmacologica Sinica |
| Volume | 35 |
| Issue | 5 |
| Page Numbers | 653-663 |
| Abstract | <strong>AIM: </strong>The chemerin receptor CMKLR1 is one type of G protein-coupled receptors abundant in monocyte-derived dendritic cells and macrophages, which plays a key role in the entry of a subset of immunodeficiency viruses including HIV/SIV into lymphocytes and macrophages. The aim of this work was to investigate how CMKLR1 was internalized and whether its internalization affected cell signaling in vitro.<br /><br /><strong>METHODS: </strong>Rat basophilic leukemia RBL-2H3 cells, HEK 293 cells, and HeLa cells were used. CMKLR1 internalization was visualized by confocal microscopy imaging or using a FACScan flow cytometer. Six potential phosphorylation sites (Ser337, Ser343, Thr352, Ser344, Ser347, and Ser350) in CMKLR1 were substituted with alanine using site-directed mutagenesis. Heterologous expression of wild type and mutant CMKLR1 allowed for functional characterization of endocytosis, Ca(2+) flux and extracellular signal-regulated kinase (ERK) phosphorylation.<br /><br /><strong>RESULTS: </strong>Chemerin and the chemerin-derived nonapeptide (C9) induced dose-dependent loss of cell surface CMKLR1-GFP fusion protein and increased its intracellular accumulation in HEK 293 cells and RBL-2H3 cells stably expressing CMKLR1. Up to 90% of CMKLR1 was internalized after treatment with C9 (1 μmol/L). By using different agents, it was demonstrated that clathrin-independent mechanism was involved in CMKLR1 internalization. Mutations in Ser343 for G protein-coupled receptor kinase phosphorylation and in Ser347 for PKC phosphorylation abrogated CMKLR1 internalization. Loss of CMKLR1 internalization partially enhanced the receptor signaling, as shown by increased Ca(2+) flux and a shorter latency to peak level of ERK phosphorylation.<br /><br /><strong>CONCLUSION: </strong>CMKLR1 internalization occurs in a clathrin-independent manner, which negatively regulated the receptor-mediated Ca(2+) flux and ERK phosphorylation. |
| Doi | 10.1038/aps.2013.198 |
| Pmid | 24658352 |
| Is Certified Translation | No |
| Dupe Override | No |
| Is Public | Yes |
| Language Text | English |