Multiple ligand recognition sites in free fatty acid receptor 2 (FFA2R) direct distinct neutrophil activation patterns

Lind, Simon; Holdfeldt, André; Mårtensson, Jonas; Granberg, Kenneth L.; Forsman, Huamei; Dahlgren, Cláes

doi:10.1016/j.bcp.2021.114762

Cited by 10 publications

(33 citation statements)

References 36 publications

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“…In order for propionate, an orthosteric FFA2R agonist, to activate the neutrophil electron transporting NADPH-oxidase system that produces superoxide anions (O 2 - ), the receptor must be allosterically modulated (see (9) and (27)). In agreement with the receptor-selectivity of propionate, the response induced by this agonist in the presence of either of the two allosteric modulators Cmp58 and AZ1729 (Fig 2), was inhibited by an FFA2R antagonist; for comparison the response was not affected by an antagonist specific for P2Y 2 R, the neutrophil receptor for ATP (Fig 3A and B).…”

Section: Resultsmentioning

confidence: 99%

“…In agreement with the receptor-selectivity of propionate, the response induced by this agonist in the presence of either of the two allosteric modulators Cmp58 and AZ1729 (Fig 2), was inhibited by an FFA2R antagonist; for comparison the response was not affected by an antagonist specific for P2Y 2 R, the neutrophil receptor for ATP (Fig 3A and B). The allosteric modulators Cmp58 and AZ1729 have been shown to be specific for FFA2R (22, 28) and to be recognized by two different allosteric receptor sites (9, 11). The response induced by propionate was not affected by the Gα q inhibitor YM-254890 (Fig 3B), showing that no Gα q containing G protein is involved in the down-stream signaling of propionate activated FFA2Rs.…”

Section: Resultsmentioning

confidence: 99%

“…The FPR2 specific hexapeptide Trp-Tyr-Met-Val-Met-NH2 (WKYMVM) was synthesized and purified by HPLC by Alta Bioscience (University of Birmingham, Birmingham, United Kingdom). The allosteric FFA2R modulator AZ1729 (9, 11, 22) together with all the other FFA2R ligands included in the study (Fig 2) were provided by AstraZeneca (Gothenburg, Sweden).…”

Section: Methodsmentioning

confidence: 99%

“…We have previously shown that the positive allosteric FFA2R modulators Cmp58 and AZ1729 are recognized by two distinct different allosteric receptor sites (8, 9). These modulators transfer propionate to a potent neutrophil activating agonist.…”

Section: Introductionmentioning

confidence: 99%

“…We recently characterized several structurally diverse compounds that, based on their positive modulating effects on the neutrophil response induced by propionate, were classified as allosteric FFA2R modulators (9). These allosteric modulators interact with either of two distinctly different allosteric modulation sites in FFA2R and we herein used these as molecular tools to determine the link (if any) between activation by the orthosteric agonist propionate and by the receptor transactivation signals generated on the inside of the plasma by cross-talking GPCRs.…”

Section: Introductionmentioning

confidence: 99%

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Signals Generated by Neutrophil Receptors for Danger Molecules Transactivate Allosterically Modulated FFA2R: Distinct response patterns are mediated by modulators recognized by different allosteric receptor sites

Lind

Granberg

Forsman

et al. 2022

Preprint

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Positive allosteric modulators for free fatty acid receptor 2 (FFA2R/GPR43), that affect receptor function through binding to two distinct allosteric binding sites, were used to determine the correlation between the responses induced in neutrophils by two distinct activation modes; FFA2R was activated either by the orthosteric agonist propionate or by a receptor transactivation mechanism that activated FFA2R from the cytosolic side of the neutrophil plasma membrane by signals generated by the neutrophil PAFR (receptor for platelet activating factor), P2Y2R (receptor for ATP), FPR1 (receptor for fMLF) and FPR2 (receptor for WKYMVM). We show that the transactivation signals that activate FFA2R in the absence of any orthosteric agonist were generated downstream of the signaling G protein that couple to PAFR and P2Y2R. This transactivation of allosterically modulated FFA2Rs, by signals generated by PAFR/P2Y2R, represents a novel mechanism by which a G protein coupled receptor can be activated. Weak correlations were obtained when the FFA2R activity was induced by the transactivation signals generated by PAFRs and P2Y2Rs were compared with the FFA2R activity induced by the orthosteric agonist propionate. Comparison of the responses for each allosteric modulator revealed that the ratio values, calculated from the peak values of the ATP and propionate responses, varied from 0.2 to 1. Depending on the allosteric modulator, the response induced by the two different mechanisms (orthosteric activation and receptor transactivation, respectively), was equal or the propionate response was more pronounced. Importantly, we conclude that FFA2R activation from outside (orthosteric activation) and inside (receptor cross-talk/transactivation) can be selectively affected by an allosteric FFA2R modulator.The allosterically modulated FFA2R is transactivated by signals generated by other GPCRs.The PAF and ATP receptors transactivate FFA2R from the cytosolic side of the membrane.The mechanisms that regulates activation of FFA2R from outside and inside differ.

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Section: Resultsmentioning

confidence: 99%