Related Experiment Video

Updated: Feb 20, 2026

Measuring the Rate of Lipolysis in Ex Vivo Murine Adipose Tissue and Primary Preadipocytes Differentiated In Vitro

Measuring the Rate of Lipolysis in Ex Vivo Murine Adipose Tissue and Primary Preadipocytes Differentiated In Vitro

Published on: March 17, 2023

Free fatty acid receptor 2 allosterism is defined by cellular context.

Simon Lind¹, Ayaan Abdi Ali², Sarah Al Hamoud Al Asswad²

¹Department of Medical Biochemistry and Cell biology, Institute of Biomedicine, Gothenburg University, Gothenburg, Sweden. Simon.lind@gu.se.

Cell Communication and Signaling : CCS

|February 19, 2026

Summary

Allosteric modulators Cmp58 and AZ1729 fine-tune G protein-coupled receptor FFA2R signaling. These modulators enable reactive oxygen species (ROS) generation in neutrophils, highlighting pathway-selective allosterism for therapeutic strategies.

Area of Science:

Pharmacology
Cellular Signaling
GPCRs

Background:

Allosteric modulators fine-tune G protein-coupled receptor (GPCR) signaling.
The short-chain fatty acid receptor FFA2R (GPR43) binds propionate and allosteric ligands like Cmp58 and AZ1729.
FFA2R signaling and allosteric modulation were characterized across HEK293, HL60, and primary human neutrophil models.

Purpose of the Study:

To characterize FFA2R signaling pathways.
To investigate the allosteric modulation of FFA2R by Cmp58 and AZ1729.
To define pathway-selective allosterism at FFA2R and identify informative readouts for therapeutic strategies.

Main Methods:

FFA2R activation assessed using G protein activation and β-arrestin recruitment assays (ebBRET biosensors).
Calcium (Ca²⁺) mobilization measured by Fura-2; reactive oxygen species (ROS) generation quantified by isoluminol chemiluminescence.
Pharmacological tools included FFA2R antagonist CATPB, Gα_q inhibitor YM-254,890, and pertussis toxin (PTX).

Main Results:

Propionate activated multiple G proteins and recruited β-arrestins in HEK293 cells.
Allosteric ligands Cmp58 and AZ1729 acted as pathway-selective ago-PAMs, potentiating Gα_i1 while attenuating Gα_q/11 and Gαi_2/3 signaling.
In neutrophils, Cmp58 or AZ1729 enabled propionate-induced ROS generation, with combined application yielding robust ROS.

Conclusions:

Calcium mobilization is a conserved FFA2R output.
Allosteric modulation by Cmp58 and AZ1729 promotes Gα_i/o-biased signaling, enabling ROS generation.
Pathway-selective allosterism at FFA2R was defined, highlighting Ca²⁺ mobilization and ROS as key readouts for therapeutic development.

Keywords:

Allosteric activation Allosteric modulation Ca2+ mobilization FFA2R Functional selectivity GPR43 HEK293 HL60 Neutrophils Reactive oxygen species

More Related Videos

A Kinetic Fluorescence-based Ca2+ Mobilization Assay to Identify G Protein-coupled Receptor Agonists, Antagonists, and Allosteric Modulators

A Kinetic Fluorescence-based Ca2+ Mobilization Assay to Identify G Protein-coupled Receptor Agonists, Antagonists, and Allosteric Modulators

Published on: February 20, 2018

Visualizing the Conformational Dynamics of Membrane Receptors Using Single-Molecule FRET

Visualizing the Conformational Dynamics of Membrane Receptors Using Single-Molecule FRET

Published on: August 17, 2022

Related Concept Videos

Allosteric Regulation

Allosteric Regulation

Allosteric regulation of enzymes occurs when the binding of an effector molecule to a site that is different from the active site causes a change in the enzymatic activity. This alternate site is called an allosteric site, and an enzyme can contain more than one of these sites. Allosteric regulation can either be positive or negative, resulting in an increase or decrease in enzyme activity. Most enzymes that display allosteric regulation are metabolic enzymes involved in the degradation or...

Allosteric Proteins-ATCase

Allosteric Proteins-ATCase

Binding sites linkages can regulate a protein's function. For example, enzyme activity is often regulated through a feedback mechanism where the end product of the biochemical process serves as an inhibitor.
Aspartate transcarbamoylase (ATCase) is a cytosolic enzyme that catalyzes the condensation of L-aspartate and carbamoyl phosphate to N-carbamoyl-L-aspartate. This reaction is the first step in pyrimidine biosynthesis. UTP and CTP, the end products of the pyrimidine synthesis...

Ligand Binding and Linkage

Ligand Binding and Linkage

Allosteric proteins have more than one ligand binding site; the binding of a ligand to any of these sites influences the binding of ligands to the other sites. When a protein is allosteric, its binding sites are called coupled or linked. In the case of enzymes, the site that binds to the substrate is known as the active site and the other site is known as the regulatory site. When a ligand binds to the regulatory site, this leads to conformational changes in the protein that can influence...

Cooperative Allosteric Transitions

Cooperative Allosteric Transitions

Cooperative allosteric transitions can occur in multimeric proteins, where each subunit of the protein has its own ligand-binding site. When a ligand binds to any of these subunits, it triggers a conformational change that affects the binding sites in the other subunits; this can change the affinity of the other sites for their respective ligands. The ability of the protein to change the shape of its binding site is attributed to the presence of a mix of flexible and stable segments in the...

Cooperative Allosteric Transitions

Cooperative Allosteric Transitions

The Two-State Receptor Model

The Two-State Receptor Model

The two-state receptor model explains a drug's interaction with receptors, such as G protein-coupled receptors and ligand-gated ion channels, to induce or inhibit a biological response. When no natural ligands are present, a receptor exists in an equilibrium of inactive (Ri) and active (Ra) conformations. The inactive form does not produce a response, while the active form generates a basal effect known as constitutive activity.
The binding affinity of a drug determines its interaction with...

Articles linked to this work by shared authors, journal, and citation graph.

Mechanistically resolved prediction of compound hepatotoxicity using primary human liver spheroids-Application to recent real-world cases.

Drug metabolism and disposition: the biological fate of chemicals·2026

GLP-1R and GIPR crosstalk modulates insulinotropic signaling pathways.

Cell chemical biology·2026

Prediction of drug hypersensitivity by comprehensive modeling of HLA-peptidomes.

Briefings in bioinformatics·2026

A Machine Vision-Guided Microphysiological Platform With Automated Microfluidics Enables Longitudinal Biomarker Monitoring and Emulation of Translationally Relevant Exposure Scenarios.

Advanced science (Weinheim, Baden-Wurttemberg, Germany)·2026

Structure-based screening and a conformational biosensor identify a GPR183 inverse agonist and an activation switch.

Nature communications·2026

Feed-Forward Deep Neural Networks Predict Substrate-Specific Effects of Transporter Variants to Explain Drug Response Variability.

Clinical and translational science·2026

See all related articles