One major bottleneck for biophysical characterization and high-resolution structure determination of G protein-coupled receptors (GPCRs) is the production of high levels of recombinant receptor. Our research has been motivated by this limitation and we are exploring the use of receptor-receptor chimeras to help improve expression and trafficking of “difficult-to-produce” receptors in yeast. The C-terminus of “well-expressed” receptors have been shown to contain important motifs necessary for efficient endoplasmic reticulum trafficking of the receptor. Here, we have utilized the C-terminus of rat tachykinin-2 receptor (NK2R) to improve the expression and trafficking of the human homologue. This human/rat chimera consists of the seven-transmembrane domains and extra- and intra-cellular loops of the human receptor and the C-terminus from the rat NK2R. Here we present results of improved expression and fluorescent ligand binding in yeast, and downstream signaling responses in yeast and HEK cells. Both the wild-type and chimeric NK2R were able to produce native-like ligand-mediated downstream signaling responses in yeast via an MAPK response pathway. The observation of native-like ligand binding and downstream signaling responses of these chimeric receptors were validated in transiently transfected mammalian cells.

Improved expression of human tachykinin NK2 receptor using a receptor-receptor chimera with the rat homologue

2019 Spring National Meeting

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.