5JQH

Structure of beta2 adrenoceptor bound to carazolol and inactive-state stabilizing nanobody, Nb60

5JQH の概要

• エントリーDOI: 10.2210/pdb5jqh/pdb
• 分子名称: Endolysin,Beta-2 adrenergic receptor, Nanobody60, Nb60, (2S)-1-(9H-Carbazol-4-yloxy)-3-(isopropylamino)propan-2-ol, ... (4 entities in total)
• 機能のキーワード: gpcr, signaling, nanobody, allostery, signaling protein, hydrolase, hydrolase-inhibitor complex, hydrolase/inhibitor
• 由来する生物種: Enterobacteria phage T4; 詳細
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 135830.68

構造登録者

Staus, D.P.,Strachan, R.T.,Manglik, A.,Pani, B.,Kahsai, A.W.,Kim, T.H.,Wingler, L.M.,Ahn, S.,Chatterjee, A.,Masoudi, A.,Kruse, A.C.,Pardon, E.,Steyaert, J.,Weis, W.I.,Prosser, R.S.,Kobilka, B.K.,Costa, T.,Lefkowitz, R.J. (登録日: 2016-05-05, 公開日: 2016-07-13, 最終更新日: 2024-10-23)

主引用文献

Staus, D.P.,Strachan, R.T.,Manglik, A.,Pani, B.,Kahsai, A.W.,Kim, T.H.,Wingler, L.M.,Ahn, S.,Chatterjee, A.,Masoudi, A.,Kruse, A.C.,Pardon, E.,Steyaert, J.,Weis, W.I.,Prosser, R.S.,Kobilka, B.K.,Costa, T.,Lefkowitz, R.J.
Allosteric nanobodies reveal the dynamic range and diverse mechanisms of G-protein-coupled receptor activation.
Nature, 535:448-452, 2016

PubMed Abstract: G-protein-coupled receptors (GPCRs) modulate many physiological processes by transducing a variety of extracellular cues into intracellular responses. Ligand binding to an extracellular orthosteric pocket propagates conformational change to the receptor cytosolic region to promote binding and activation of downstream signalling effectors such as G proteins and β-arrestins. It is well known that different agonists can share the same binding pocket but evoke unique receptor conformations leading to a wide range of downstream responses (‘efficacy’). Furthermore, increasing biophysical evidence, primarily using the β2-adrenergic receptor (β2AR) as a model system, supports the existence of multiple active and inactive conformational states. However, how agonists with varying efficacy modulate these receptor states to initiate cellular responses is not well understood. Here we report stabilization of two distinct β2AR conformations using single domain camelid antibodies (nanobodies)—a previously described positive allosteric nanobody (Nb80) and a newly identified negative allosteric nanobody (Nb60). We show that Nb60 stabilizes a previously unappreciated low-affinity receptor state which corresponds to one of two inactive receptor conformations as delineated by X-ray crystallography and NMR spectroscopy. We find that the agonist isoprenaline has a 15,000-fold higher affinity for β2AR in the presence of Nb80 compared to the affinity of isoprenaline for β2AR in the presence of Nb60, highlighting the full allosteric range of a GPCR. Assessing the binding of 17 ligands of varying efficacy to the β2AR in the absence and presence of Nb60 or Nb80 reveals large ligand-specific effects that can only be explained using an allosteric model which assumes equilibrium amongst at least three receptor states. Agonists generally exert efficacy by stabilizing the active Nb80-stabilized receptor state (R80). In contrast, for a number of partial agonists, both stabilization of R80 and destabilization of the inactive, Nb60-bound state (R60) contribute to their ability to modulate receptor activation. These data demonstrate that ligands can initiate a wide range of cellular responses by differentially stabilizing multiple receptor states.

PubMed: 27409812
DOI: 10.1038/nature18636

実験手法

X-RAY DIFFRACTION (3.2 Å)