GPCR Engineering Yields High-Resolution Structural Insights into β2-Adrenergic Receptor Function

@article{Rosenbaum2007GPCREY,
  title={GPCR Engineering Yields High-Resolution Structural Insights into $\beta$2-Adrenergic Receptor Function},
  author={Daniel M. Rosenbaum and Vadim Cherezov and Michael A. Hanson and S{\o}ren G. F. Rasmussen and Foon Sun Thian and Tong Sun Kobilka and Hee-Jung Choi and Xiao Jie Yao and William I. Weis and Raymond C. Stevens and Brian K. Kobilka},
  journal={Science},
  year={2007},
  volume={318},
  pages={1266 - 1273}
}
The β2-adrenergic receptor (β2AR) is a well-studied prototype for heterotrimeric guanine nucleotide–binding protein (G protein)–coupled receptors (GPCRs) that respond to diffusible hormones and neurotransmitters. To overcome the structural flexibility of the β2AR and to facilitate its crystallization, we engineered a β2AR fusion protein in which T4 lysozyme (T4L) replaces most of the third intracellular loop of the GPCR (“β2AR-T4L”) and showed that this protein retains near-native pharmacologic… 
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Structural equilibrium underlying ligand-dependent activation of β2-adrenoreceptor
TLDR
NMR structural analysis of an active state of the β2-adrenergic receptor defines a unique orientation for the intracellular half of TM6, responsible for G-protein binding, including an equilibrium among three conformations of a key microswitch.
Structure and Function of an Irreversible Agonist-β2 Adrenoceptor complex
TLDR
A covalent agonist-bound β2AR–T4L fusion protein is designed that can be covalently tethered to a specific site on the receptor through a disulphide bond, and is capable of activating a heterotrimeric G protein.
Crystal Structure of the β2Adrenergic Receptor-Gs protein complex
TLDR
This crystal structure represents the first high-resolution view of transmembrane signalling by a GPCR and the most surprising observation is a major displacement of the α-helical domain of Gαs relative to the Ras-like GTPase domain.
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References

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High-Resolution Crystal Structure of an Engineered Human β2-Adrenergic G Protein–Coupled Receptor
TLDR
Although the location of carazolol in the β2-adrenergic receptor is very similar to that of retinal in rhodopsin, structural differences in the ligand-binding site and other regions highlight the challenges in using rhodopin as a template model for this large receptor family.
Coupling ligand structure to specific conformational switches in the β2-adrenoceptor
TLDR
It is found that most partial agonists were as effective as full agonists in disrupting the ionic lock, and disruption of this important molecular switch is necessary, but not sufficient, for full activation of the β2-AR.
Crystal structure of the human β2 adrenergic G-protein-coupled receptor
TLDR
The β2AR structure differs from rhodopsin in having weaker interactions between the cytoplasmic ends of transmembrane (TM)3 and TM6, involving the conserved E/DRY sequences, which may be responsible for the relatively high basal activity and structural instability of the β2 AR.
Agonist-induced conformational changes in the G-protein-coupling domain of the β2 adrenergic receptor
TLDR
The authors' studies, when compared with studies of activation in rhodopsin, indicate a general mechanism for GPCR activation; however, a notable difference is the relatively slow kinetics of the conformational changes in the β2AR, which may reflect the different energetics of activation by diffusible ligands.
Probing the β2 Adrenoceptor Binding Site with Catechol Reveals Differences in Binding and Activation by Agonists and Partial Agonists*
TLDR
Catechol is used as a molecular probe to identify mechanistic differences between β2AR activation by catecholamine agonists, such as isoproterenol, and by the structurally related non-catechol partial agonist salbutamol, showing unexpected differences in binding and activation by structurally similar agonists and partial agonists.
Agonists induce conformational changes in transmembrane domains III and VI of the β2 adrenoceptor
TLDR
Findings suggest that IANBD bound to 125Cys and 285Cys are exposed to a more polar environment upon agonist binding, and indicate that movements of transmembrane segments III and VI are involved in activation of G protein‐coupled receptors.
Functionally Different Agonists Induce Distinct Conformations in the G Protein Coupling Domain of the β2Adrenergic Receptor*
TLDR
Monitoring ligand-induced conformational changes in the G protein-coupling domain of the β2 adrenergic receptor provides new insight into the structural consequence of antagonist binding and the basis of agonism and partial agonism.
Sequential binding of agonists to the beta2 adrenoceptor. Kinetic evidence for intermediate conformational states.
TLDR
A mechanistic model for GPCR activation where contacts between the receptor and structural determinants of the agonist stabilize a succession of conformational states with distinct cellular functions is supported.
Role of group-conserved residues in the helical core of β2-adrenergic receptor
TLDR
To determine the role of group-conserved residues in the β2-adrenergic receptor (β2-AR), amino acid replacements guided by molecular modeling were carried out at key positions in transmembrane helices H2–H4 and allow insights into the roles of these residues in GPCR structure and function.
Structural Instability of a Constitutively Active G Protein-coupled Receptor
TLDR
It is proposed that the mutation that confers constitutive activity to the β2 adrenergic receptor removes some stabilizing conformational constraints, allowing CAM to more readily undergo transitions between the inactive and the active states and making the receptor more susceptible to denaturation.
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