Targeting G protein-coupled receptor signaling at the G protein level with a selective nanobody inhibitor.

Gulati, Sahil; Jin, Hui; Masuho, Ikuo; Orban, Tivadar; Cai, Yuan; Pardon, Els; Martemyanov, Kirill A; Kiser, Philip D et al. · Nat Commun · 2018

basic_science · Level V

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Abstract

G protein-coupled receptors (GPCRs) activate heterotrimeric G proteins by mediating a GDP to GTP exchange in the Gα subunit. This leads to dissociation of the heterotrimer into Gα-GTP and Gβγ dimer. The Gα-GTP and Gβγ dimer each regulate a variety of downstream pathways to control various aspects of human physiology. Dysregulated Gβγ-signaling is a central element of various neurological and cancer-related anomalies. However, Gβγ also serves as a negative regulator of Gα that is essential for G protein inactivation, and thus has the potential for numerous side effects when targeted therapeutically. Here we report a llama-derived nanobody (Nb5) that binds tightly to the Gβγ dimer. Nb5 responds to all combinations of β-subtypes and γ-subtypes and competes with other Gβγ-regulatory proteins for a common binding site on the Gβγ dimer. Despite its inhibitory effect on Gβγ-mediated signaling, Nb5 has no effect on Gα<sub>q</sub>-mediated and Gα<sub>s</sub>-mediated signaling events in living cells.

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