Lysophosphatidic acid (LPA) stimulates cells by activation of five G-protein-coupled receptors, termed LPA1-5. The LPA1 receptor is the most widely expressed and is a major regulator of cell migration. In this study, we show that phorbol ester (PMA)-induced internalization of the LPA1 receptor requires clathrin AP-2 complexes, protein kinase C, and a distal dileucine motif (amino acids 352 and 353) in the cytoplasmic tail but not β-arrestin. Agonist-dependent internalization of LPA1, however, requires a cluster of serine residues (amino acids 341-347) located proximal to the dileucine motif, β-arrestin, and to a lesser extent clathrin AP-2. The serine cluster of LPA1 is required for β-arrestin2-GFP translocation to the plasma membrane and signal desensitization. In contrast, the dileucine motif (IL) is required for both basal and PMA-induced internalization. Evidence for the β-arrestin independence of PMA-induced internalization of LPA1 comes from the observations that β-arrestin2-GFP is not recruited to the plasma membrane upon PMA treatment and that LPA1 is readily internalized in β-arrestin1/2 knock-out mouse embryonic fibroblasts. These results indicate that distinct molecular mechanisms regulate agonist-dependent and PMA-dependent internalization of the LPA1 receptor.
All Science Journal Classification (ASJC) codes
- Molecular Biology
- Cell Biology