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bioRxiv · 10.64898/2026.10.26.761934

Unliganded Active State of the β2-Adrenergic Receptor Revealed by Pressure-Resolved DEER and ProGuide Molecular Modeling

Lerch, M.·Brennan, P. C.·Garces, A. M.·Grosskopf, J. D.·Pani, B.
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Abstract

The intrinsic ability of G protein-coupled receptors (GPCRs), such as the {beta}2-adrenergic receptor ({beta}2AR), to sample an ensemble of distinct conformational states with varying affinities and catalytic activities for intracellular transducers such as G proteins and {beta} arrestins is a feature that underlies their signaling complexity. The structural features of the full ensemble of accessible conformations remain largely inaccessible to conventional biophysical techniques due to the low populations of transient or intermediate states. Here, we integrate pressure-resolved double electron electron resonance (PR DEER) with the ProGuide biased molecular dynamics framework to map elusive {beta}2AR conformations obscured under ambient conditions. Our results demonstrate a pressure dependent, global two-state equilibrium between inactive- and active-like conformations in the unliganded receptor and indicate a high degree of allosteric coupling among transmembrane helices in the absence of ligand binding. Notably, {beta}2AR bound to the superagonist BI 167107 samples a smaller conformational space than the unliganded active conformation revealed by pressure, suggesting that this agonist constrains {beta}2AR to a more restricted subset of conformers within the active ensemble. This finding points towards a conformational selection mechanism of activation of the {beta}2AR wherein signaling variability exists because of conformational heterogeneity within the active state.

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Lerch, M., Brennan, P. C., Garces, A. M., Grosskopf, J. D., Pani, B.. 2026-10-27. Unliganded Active State of the β2-Adrenergic Receptor Revealed by Pressure-Resolved DEER and ProGuide Molecular Modeling. https://doi.org/10.64898/2026.10.26.761934

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