Journal
BIOCHIMICA ET BIOPHYSICA ACTA-MOLECULAR CELL RESEARCH
Volume 1867, Issue 12, Pages -Publisher
ELSEVIER
DOI: 10.1016/j.bbamcr.2020.118849
Keywords
G protein-coupled receptors; FPR2; AP2; Actin cytoskeleton; Barbadin; beta-Arrestin; Desensitization; Resensitization; Endocytosis; Neutrophils; Reactive oxygen species
Categories
Funding
- Swedish Research Council [2015-02448]
- King Gustaf V's 80-Year Foundation [FAI-2014-0011, FAI2014-0029]
- Swedish Foundation for Strategic Research [SM17-0046]
- Magnus Bergvall Foundation [MS2018-02579]
- Swedish Government under the ALF-agreement
- Rune and Ulla Almlovs Foundation (MS 2019)
- Elisabeth and Alfred Ahlqvist's foundation - Swedish Pharmacy Society Grant (MS 2020)
- Ingabritt and Arne Lundberg Research Foundation
- European Union's Horizon 2020 research and innovation programme (Marie Sklodowska-Curie grant [797497]
- Swedish Society for Medical Research [P18-0098]
- Canadian Institutes of Health Research (CIHR) [FDN-148431]
- Canada Research Chair in Signal Transduction and Molecular Pharmacology
- Vinnova [2015-02448] Funding Source: Vinnova
- Marie Curie Actions (MSCA) [797497] Funding Source: Marie Curie Actions (MSCA)
- Swedish Foundation for Strategic Research (SSF) [SM17-0046] Funding Source: Swedish Foundation for Strategic Research (SSF)
- Swedish Research Council [2015-02448] Funding Source: Swedish Research Council
Ask authors/readers for more resources
FPR2, a member of the family of G protein-coupled receptors (GPCRs), mediates neutrophil migration, a response that has been linked to beta-arrestin recruitment. beta-Arrestin regulates GPCR endocytosis and can also elicit non-canonical receptor signaling. To determine the poorly understood role of beta-arrestin in FPR2 endocytosis and in NADPH-oxidase activation in neutrophils, Barbadin was used as a research tool in this study. Barbadin has been shown to bind the clathrin adaptor protein (AP2) and thereby prevent beta-arrestin/AP2 interaction and beta-arrestin-mediated GPCR endocytosis. In agreement with this, AP2/beta-arrestin interaction induced by an FPR2-specific agonist was inhibited by Barbadin. Unexpectedly, however, Barbadin did not inhibit FPR2 endocytosis, indicating that a mechanism independent of beta-arrestin/AP2 interaction may sustain FPR2 endocytosis. This was confirmed by the fact, that FPR2 also underwent agonist-promoted endocytosis in beta-arrestin deficient cells, albeit at a diminished level as compared to wild type cells. Dissection of the Barbadin effects on FPR2-mediated neutrophil functions including NADPH-oxidase activation mediated release of reactive oxygen species (ROS) and chemotaxis revealed that Barbadin had no effect on chemotactic migration whereas the release of ROS was potentiated/primed. The effect of Barbadin on ROS production was reversible, independent of beta-arrestin recruitment, and similar to that induced by latrunculin A. Taken together, our data demonstrate that endocytic uptake of FPR2 occurs independently of beta-arrestin, while Barbadin selectively augments FPR2-mediated ROS production independently of receptor endocytosis. Given that Barbadin binds to AP2 and prevents the AP2/beta-arrestin interaction, our results indicate a role for AP2 in FPR2-mediated ROS release from neutrophils.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available