High throughput mutational characterization of the GPCR ligand C5a using yeast display and deep sequencing

As the largest class of membrane receptors and drug targets, G protein-coupled receptors (GPCRs) have been the focus of these studies. While significant progress has been made in characterizing the receptors, mutagenesis studies on their ligands have lagged behind, because of the challenges in solubilizing the target receptor.

In the publication titled "High throughput mutational characterization of the GPCR ligand C5a using yeast display and deep sequencing", we present a novel approach that utilizes lipid vesicles to embed and stabilize target membrane receptors, allowing direct ligand screening. Using this platform, we investigate the anaphylatoxin Complement 5a (C5a) and characterized the mutational impacts on its interactions with the two native GPCRs: complement 5a receptor 1 (C5aR1) and complement 5a receptor 2 (C5L2).