GPCR Ligand Profiling/Phosphorylation Barcoding Services

Creative Biolabs is the specialist provider of GPCR ligand profiling/phosphorylation barcoding services. We have a team of professional scientists to provide customers with comprehensive and rapid high-throughput GPCR ligand reporting and phosphorylation barcoding reporting services. We will screen potential GPCR ligands and provide comprehensive phosphorylation barcoding reports through standard experimental procedures to meet our clients' GPCR drug discovery needs.

GPCR ligand profiling

G protein-coupled receptor (GPCR) is the best-known drug target molecule. Therefore, high-throughput GPCR ligand analysis remains the main power of GPCR drug discovery. Currently, the ideal GPCR ligand screening method is the microtitration plate format (96-well, 384-well, or 1536-well) with no radioactivity, stability, and automated detection. In high-throughput screening, GPCR binds to ligands and the activated G_q induces Phospholipase C (PLC) production. The metabolite of PLC was inositol-1-phosphate (IP1). By the method of Homogeneous Time-Resolved Fluorescence (HTRF), free IP1 and labeled IP1 are competitively bound to antibodies. Thus, the fluorescence resonance Energy Transfer Technology (FRET) process is blocked to achieve rapid and high-throughput detection of IP1 molecular concentration in samples, thus realizing the screening of GPCRs ligands. Creative Biolabs offers a new generation of high-throughput GPCR ligand profiling, with a large selection of GPCR stable cell lines. Customers can quickly screen out ideal GPCR ligands by sending us a library of compounds of interest.

Fig.1 Receptor dimerization assays. (Cechova, 2021)

GPCR phosphorylation barcoding

The closure of the GPCR signaling pathway is determined by GPCR phosphorylation barcoding. After ligand binding to GPCR, different GPCR kinases (GRKs) can lead to different forms of phosphorylation of the C-terminal serine and threonine residues of GPCR (visualized as phosphorylation barcoding), and the phosphorylation barcoding may be biased by the conformation of the receptor and ligand. This phosphorylated barcoding recruits β-arrestins to regulate subsequent GPCR desensitization, bias signal transduction, and endocytosis. Many diseases are known to be associated with the phosphorylation of GPCR. Therefore, exploring where phosphorylation occurs at different GPCR sites plays a key role in GPCR drug discovery. Mass spectrometry (MS) is a powerful tool for sequencing phosphorylated residues of different proteins in complex protein mixtures. The phosphorylated GPCR was enriched by solid-phase metal affinity chromatography, phosphorylated antibodies, or TiO2, and during the enrichment process, the single-phosphorylated and multi-phosphorylated peptides were separated step by step, followed by mass spectrometry. Creative Biolabs provides a GPCR phosphorylation barcode sequencing service based on MS technology to report details of identified phosphorylation sites.

Fig.2. Phosphorylation barcoding controls β-arrestin conformation, leading to multiple signaling options. (Liggett, 2011)

Our Feature Service

• Gold standard service of GPCR ligand detection and GPCR phosphorylation barcoding detection.
• Providing a wide range of target GPCR stabilized cell lines.
• High throughput: large numbers of compounds and proteins can be screened at one time.
• Providing customers with comprehensive and detailed raw data and analysis reports.
• Quick turnaround times of 1-3 weeks depending on project types.

Creative Biolabs provides optimizing GPCR ligand profiling and GPCR phosphorylation barcoding services. Please contact us to discuss your project and we will tailor your high-throughput GPCR ligand profiling regimen and GPCR phosphorylated barcode analysis regimen to achieve your goals.

References

1. Trinquet, E. Monitoring Gq-coupled receptor response through inositol phosphate quantification with the IP-One assay. Expert Opinion on Drug Discovery. 2011; 6(10):981-994.
2. Liggett, S.B. Phosphorylation barcoding as a mechanism of directing GPCR signaling. Science Signaling. 2011; 4(185): pe36.