The dynamics of agonist-β₂-adrenergic receptor activation induced by binding of GDP-bound Gs protein

There is considerable uncertainty about the mechanism by which the β₂-adrenergic receptor (β₂AR) is activated. Here we use molecular metadynamics computations to predict the mechanism by which an agonist induces the activation of the β₂AR and its cognate Gs protein. We found that binding agonist alone to the inactive β₂AR does not break the ionic lock and hence does not drive the β₂AR towards the activated conformation. However, we found that attaching the inactive Gs protein to the agonist-bound inactive β₂AR (containing the ionic lock) leads to partial insertion of Gαs-α5 into the core of β₂AR, which breaks the ionic lock, leading to activation of the Gs protein coupled to β₂AR. Upon activation, the Gαs protein undergoes a remarkable opening of the GDP binding pocket, making the GDP available for exchange or release. Concomitantly, Gαs-α5 undergoes a remarkable expansion in the β₂AR cytoplasmic region after the ionic lock is broken, inducing TM6 to displace outward by ~5 Å from TM3.