- Nygaard, Rie;
- Zou, Yaozhong;
- Dror, Ron O;
- Mildorf, Thomas J;
- Arlow, Daniel H;
- Manglik, Aashish;
- Pan, Albert C;
- Liu, Corey W;
- Fung, Juan José;
- Bokoch, Michael P;
- Thian, Foon Sun;
- Kobilka, Tong Sun;
- Shaw, David E;
- Mueller, Luciano;
- Prosser, R Scott;
- Kobilka, Brian K
G-protein-coupled receptors (GPCRs) can modulate diverse signaling pathways, often in a ligand-specific manner. The full range of functionally relevant GPCR conformations is poorly understood. Here, we use NMR spectroscopy to characterize the conformational dynamics of the transmembrane core of the β(2)-adrenergic receptor (β(2)AR), a prototypical GPCR. We labeled β(2)AR with (13)CH(3)ε-methionine and obtained HSQC spectra of unliganded receptor as well as receptor bound to an inverse agonist, an agonist, and a G-protein-mimetic nanobody. These studies provide evidence for conformational states not observed in crystal structures, as well as substantial conformational heterogeneity in agonist- and inverse-agonist-bound preparations. They also show that for β(2)AR, unlike rhodopsin, an agonist alone does not stabilize a fully active conformation, suggesting that the conformational link between the agonist-binding pocket and the G-protein-coupling surface is not rigid. The observed heterogeneity may be important for β(2)AR's ability to engage multiple signaling and regulatory proteins.