Decision letter: Computational design of thermostabilizing point mutations for G protein-coupled receptors

Transmembranal G Protein-Coupled Receptors (GPCRs) transduce extracellular chemical signals to the cell, via conformational change from a resting (inactive) to an active (canonically bound to a G-protein) conformation. Receptor activation is normally modulated by extracellular ligand binding, but mutations in the receptor can also shift this equilibrium by stabilizing different conformational states. In this work, we built structure-energetic relationships of receptor activation based on original thermodynamic cycles that represent the conformational equilibrium of the prototypical A2A adenosine receptor (AR). These cycles were solved with efficient free energy perturbation (FEP) protocols, allowing to distinguish the pharmacological profile of different series of A2AAR agonists with different efficacies. The modulatory effects of point mutations on the basal activity of the receptor or on ligand efficacies could also be detected. This methodology can guide GPCR ligand design with tailored pharmacological properties, or allow the identification of mutations that modulate receptor activation with potential clinical implications.

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Stabilization of G protein-coupled receptors by point mutations

Frontiers in Pharmacology ◽

10.3389/fphar.2015.00082 ◽

2015 ◽

Vol 6 ◽

Cited By ~ 36

Author(s):

Franziska M. Heydenreich ◽

Ziva Vuckovic ◽

Milos Matkovic ◽

Dmitry B. Veprintsev

Keyword(s):

G Protein ◽

Point Mutations ◽

G Protein Coupled Receptors ◽

G Protein Coupled

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Computational design of thermostabilizing point mutations for G protein-coupled receptors

eLife ◽

10.7554/elife.34729 ◽

2018 ◽

Vol 7 ◽

Cited By ~ 29

Author(s):

Petr Popov ◽

Yao Peng ◽

Ling Shen ◽

Raymond C Stevens ◽

Vadim Cherezov ◽

...

Keyword(s):

Structural Information ◽

Transmembrane Protein ◽

Point Mutations ◽

Computational Design ◽

G Protein Coupled Receptors ◽

Single Mutation ◽

Triple Mutant ◽

Receptor Complexes ◽

Therapeutic Drugs ◽

G Protein Coupled

Engineering of GPCR constructs with improved thermostability is a key for successful structural and biochemical studies of this transmembrane protein family, targeted by 40% of all therapeutic drugs. Here we introduce a comprehensive computational approach to effective prediction of stabilizing mutations in GPCRs, named CompoMug, which employs sequence-based analysis, structural information, and a derived machine learning predictor. Tested experimentally on the serotonin 5-HT2C receptor target, CompoMug predictions resulted in 10 new stabilizing mutations, with an apparent thermostability gain ~8.8°C for the best single mutation and ~13°C for a triple mutant. Binding of antagonists confers further stabilization for the triple mutant receptor, with total gains of ~21°C as compared to wild type apo 5-HT2C. The predicted mutations enabled crystallization and structure determination for the 5-HT2C receptor complexes in inactive and active-like states. While CompoMug already shows high 25% hit rate and utility in GPCR structural studies, further improvements are expected with accumulation of structural and mutation data.

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Deciphering conformational selectivity in the A2A adenosine G protein-coupled receptor by free energy simulations

PLoS Computational Biology ◽

10.1371/journal.pcbi.1009152 ◽

2021 ◽

Vol 17 (11) ◽

pp. e1009152

Author(s):

Willem Jespers ◽

Laura H. Heitman ◽

Adriaan P. IJzerman ◽

Eddy Sotelo ◽

Gerard J. P. van Westen ◽

...

Keyword(s):

Free Energy ◽

G Protein ◽

Point Mutations ◽

Receptor Activation ◽

G Protein Coupled Receptors ◽

Free Energy Perturbation ◽

Thermodynamic Cycles ◽

Energy Perturbation ◽

Energy Simulations ◽

G Protein Coupled

Transmembranal G Protein-Coupled Receptors (GPCRs) transduce extracellular chemical signals to the cell, via conformational change from a resting (inactive) to an active (canonically bound to a G-protein) conformation. Receptor activation is normally modulated by extracellular ligand binding, but mutations in the receptor can also shift this equilibrium by stabilizing different conformational states. In this work, we built structure-energetic relationships of receptor activation based on original thermodynamic cycles that represent the conformational equilibrium of the prototypical A2A adenosine receptor (AR). These cycles were solved with efficient free energy perturbation (FEP) protocols, allowing to distinguish the pharmacological profile of different series of A2AAR agonists with different efficacies. The modulatory effects of point mutations on the basal activity of the receptor or on ligand efficacies could also be detected. This methodology can guide GPCR ligand design with tailored pharmacological properties, or allow the identification of mutations that modulate receptor activation with potential clinical implications.

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