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Rescue of internalization-defective platelet-activating factor receptor function by EBP50/NHERF1

Journal of Cell Communication and Signaling volume 6, pages 205–216 (2012)Cite this article

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.

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Abbreviations

CFTR:

Cystic fibrosis transmembrane conductance regulator

EBP50:

Ezrin-radixin-moesin binding protein of 50 kDa

GPCR:

G-protein coupled receptor

GRK:

GPCR-specific kinases

NHERF1:

Na+/H+ exchanger regulatory factor 1

PAF:

Platelet-activating factor

WT:

Wild-type

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Acknowledgements

This work was supported by grants (MRP, JS) from the Canadian Institutes for Health Research and from the Natural Sciences and Engineering Research Council of Canada (NSERC RGPIN-355310-2008) (DJD). DJD holds a CIHR New Investigator Salary Award.

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Affiliations

  1. Department of Pharmacology, Faculty of Medicine, Dalhousie University, 5850 College St., PO BOX 15000, Halifax, NS, B3H 4R2, Canada

    Denis J. Dupré

  2. Department of Pediatrics, Immunology Division, Faculty of Medicine, Université de Sherbrooke, 3001 12th Avenue North, Sherbrooke, QC, J1H 5N4, Canada

    Marek Rola-Pleszczynski & Jana Stankova

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  1. Denis J. Dupré
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  2. Marek Rola-Pleszczynski
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  3. Jana Stankova
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Correspondence to Denis J. Dupré or Jana Stankova.

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Concise 2–3 sentence summary

EBP50 has previously been shown to regulate several functions of G protein coupled receptors. Here, we show that EBP50 regulates several functions of the platelet-activating factor receptor, and acts in concert with arrestins and GRKs to regulate receptor internalization.

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Dupré, D.J., Rola-Pleszczynski, M. & Stankova, J. Rescue of internalization-defective platelet-activating factor receptor function by EBP50/NHERF1. J. Cell Commun. Signal. 6, 205–216 (2012). https://doi.org/10.1007/s12079-012-0175-1

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Keywords

  • Arrestin
  • EBP50/NHERF1
  • GPCR
  • GRK
  • Internalization
  • PAFR