Analysis of full and partial agonists binding to β2‐adrenergic receptor suggests a role of transmembrane helix V in agonist‐specific conformational changes

The 2.4 A crystal structure of the β2-adrenergic receptor (β2AR) in complex with the high-affinity inverse agonist (-)-carazolol provides a detailed structural framework for the analysis of ligand recognition by adrenergic receptors. Insights into agonist binding and the corresponding conformational changes triggering GPCR activation mechanism are of special interest. Here we show that while the carazolol pocket captured in the β2AR crystal structure accommodates (-)-isoproterenol and other agonists without steric clashes, a finite movement of the flexible extracellular part of TMV helix (TM-Ve) obtained by receptor optimization in the presence of docked ligand can further improve the calculated binding affinities for agonist compounds. Tilting of TM-Ve towards the receptor axis provides a more complete description of polar receptor/ligand interactions for full and partial agonists, by enabling optimal engagement of agonists with two experimentally identified anchor sites, formed by Asp113/Asn312 and Ser203/Ser204/Ser207 side chains. Further, receptor models incorporating a flexible TM-V backbone allow reliable prediction of binding affinities for a set of diverse ligands, suggesting potential utility of this approach to design of effective and subtypespecific agonists for adrenergic receptors. Systematic differences in capacity of partial, full and inverse agonists to induce TM-V helix tilt in the β2AR model suggest potential role of TM-V as a conformational “rheostat” involved in the whole spectrum of β2AR responses to small molecule signals.