title: Rescue of internalization-defective platelet-activating factor receptor function by EBP50/NHERF1.
abstract: Platelet-activating factor (PAF) is a potent phospholipid mediator involved in specific disease states such as allergic asthma, atherosclerosis and psoriasis. The human PAF receptor (PAFR) is a member of the G protein-coupled receptor (GPCR) family. Following PAF stimulation, cells become rapidly desensitized; this refractory state can be maintained for hours and is dependent on PAFR phosphorylation, internalization and trafficking. EBP50/NHERF1 has been found to interact with a variety of proteins and these interactions are involved in a growing range of functions including the assembly of signalling complexes, receptor recycling and transport of proteins to the cell surface. Crucial roles of EBP50 in GPCR physiology include its involvement in internalization, recycling, and downregulation. We were interested in identifying the role of EBP50 in PAFR trafficking. Our results showed that EBP50 binds the PAFR in its basal state, while stimulation decreased the ratio of interaction between the two proteins. We also demonstrated that EBP50 could bind PAFR via its PDZ 2 domain. In addition, we studied the role of EBP50 in various functions of the PAFR such as PAF-induced inositol phosphate accumulation and receptor internalization: EBP50 decreased the WT PAFR response and rescued the function of internalization-deficient mutant receptors, as previously described for the arrestins and the GRKs. These results suggest new roles for EBP50, some of which could help understanding the complex formation after receptor activation.
authors.affiliation: Department of Pharmacology, Faculty of Medicine, Dalhousie University, 5850 College St., PO BOX 15000, Halifax, NS, B3H 4R2, Canada, denis.dupre@dal.ca.
authors.forename: Denis J
authors.initials: DJ
authors.lastname: Dupré
authors.forename: Marek
authors.initials: M
authors.lastname: Rola-Pleszczynski
authors.forename: Jana
authors.initials: J
authors.lastname: Stankova