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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Structure and Function of an Irreversible Agonist-β2 Adrenoceptor complex

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References

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High-Resolution Crystal Structure of an Engineered Human β2-Adrenergic G Protein–Coupled Receptor

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

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.

Agonist-induced conformational changes in the G-protein-coupling domain of the β2 adrenergic receptor

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.

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Agonists induce conformational changes in transmembrane domains III and VI of the β2 adrenoceptor

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*

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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.

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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

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.

A dileucine motif in the C terminus of the beta2-adrenergic receptor is involved in receptor internalization.

Evidence is provided for an essential role of this motif in the agonist-induced internalization of the beta2-adrenergic receptor in the L339,340A double mutant and reduced in the two single mutants.
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