GPCR Assay Kits
Creative Biolabs has the assays you can rely on for high throughput screening, lead optimization, characterizing and discovering targets, and uncovering the complexity of disease pathways. We can offer membrane protein in vitro assay kits that save valuable laboratory time and is ideal for high throughput screening.
Background of GPCR
The most numerous and varied class of membrane receptors found in eukaryotes are G-protein-coupled receptors (GPCRs). For communications in the form of light energy, peptides, lipids, carbohydrates, and proteins, these cell surface receptors function as an inbox. GPCRs are involved in a staggering variety of bodily processes, and better knowledge of these receptors has had a significant impact on modern medicine. In fact, according to research, between one-third and fifty percent of all commercially available medications work by attaching to GPCRs.
Features of GPCR Assay Kits
To meet the various requirements of your drug development programs, Creative Biolabs offers a variety of GPCR assay kits. These detection reagents are designed to be used in a wide range of applications, including second messenger assays, translocation, compound-protein binding, and cell-based protein-protein interactions.
- Detection of β-arrestin, calcium, and cAMP using no-wash assays
- Compatible with higher density plate formats (96-, 384-, etc.)
- Comprehensive assay procedures from cells to assays
- Completion solution developed and supported by Creative Biolabs' experts
Published Data
| Paper Title | Conformational selection guides β-arrestin recruitment at a biased G protein-coupled receptor |
| Journal | Science |
| Published | 2022 |
| Abstract | To coordinate a wide range of cellular processes, G protein-coupled receptors, also known as GPCRs, recruit β-arrestins; however, the structural dynamics underlying this process are poorly understood. Atypical chemokine receptors (ACKRs), also referred to as GPCRs, are inherently biased and only interact with β-arrestins rather than G proteins. As a result, they are used as a model system to study the structural underpinnings of β-arrestin recruitment. ACKR3 that had been labeled with 13CH3-ε-methionine was used in experiments using nuclear magnetic resonance (NMR), and the outcomes demonstrated that β-arrestin recruitment is connected to conformational exchange at crucial regions of the extracellular ligand-binding pocket and the intracellular β-arrestin-coupling region. The identification of an allosteric hub in the receptor core was made possible by NMR analyses of ACKR3 mutants with impaired β-arrestin recruitment. This hub orchestrates changes between conformational states that are both selectively populated and heterogeneously populated. Their results suggest that by adjusting receptor dynamics at intracellular and extracellular locations, conformational selection is responsible for directing the recruitment of β-arrestin. |
| Result |
In accordance with previous research, ACKR3 recruits β-arrestin without activating G protein signaling. To investigate the mechanisms underlying β-arrestin recruitment at ACKR3, β-arrestin-2 recruitment was measured in response to a panel of ACKR3 agonists, including the endogenous chemokine CXCL12 (EC50= 0.75 nM), the small molecule CCX777 (EC50 = 0.95 nM; Emax (% of CXCL12) = 75 ± 2%), and a recently described peptide LIH383. Researchers also found a powerful, extracellularly-targeting ACKR3 competitive antagonis called VUN701 (IC50 = 1.47 uM). These four ligands demonstrate a spectrum of β-arrestin-2 recruitment activities, allowing us to sample inactive (i.e., VUN701-bound) and active (i.e., CCX777-, LIH383-, and CXCL12-bound) β-arrestin recruiting states of ACKR3.
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Fig.1 β-arrestin-biased signaling at ACKR3. (Kleist, 2022)
Reference
- Kleist, A.B.; et al. Conformational selection guides β-arrestin recruitment at biased G protein-coupled receptor. Science. 2022, 377(6602): 222—228.
