Hiroki Nakai, Kinuka Isshiki, Masato Hattori, Hiromasa Maehira, Tatsumi Yamaguchi, Keiko Masuda, Yoshihiro Shimizu, Takayoshi Watanabe, Takahiro Hohsaka, Wataru Shihoya, Osamu Nureki, Yasuhiko Kato, Hajime Watanabe, and Tomoaki Matsuura
Engineering G-protein-coupled receptors (GPCRs) for improved stability or altered function is of great interest, as GPCRs consist of the largest protein family, are involved in many important signaling pathways, and thus, are one of the major drug targets. Here, we report the development of a high-throughput screening method for GPCRs using a reconstituted in vitro transcription-translation (IVTT) system. Human endothelin receptor type-B (ETBR), a class A GPCR that binds endothelin-1 (ET-1), a 21-residue peptide hormone, was synthesized in the presence of nanodisc (ND) composed of a phospholipid, 1-palmitoyl-2-oleoyl