Fine-Tuning of the Neuropeptide Y1 G Protein-Coupled Receptor by the Tryptophan6.48 “Toggle Switch”

TL;DR

This study explores how a specific amino acid in a receptor protein influences its signaling behavior, suggesting it acts as a fine-tuner rather than a main switch.

Contribution

The study reveals that tryptophan W6.48 in the Y1R receptor fine-tunes signaling rather than being a primary activation switch.

Findings

W6.48 adopts multiple rotameric states linked to backbone structure.

Mutation of W6.48 does not block G-protein signaling but reduces ligand binding efficiency.

W6.48 influences Y1R conformational similarity to active states, affecting signaling dynamics.

Abstract

G protein-coupled receptors (GPCRs) transduce extracellular
signals
into the cell through binding and activation of intracellular effector
proteins. Highly conserved residues such as tryptophan W6.48 of transmembrane helix 6 can play a role in GPCR activation, where
W6.48 acts as a microswitch, changing its rotameric state
depending on whether the receptor is bound to an agonist or antagonist.
However, its exact role is not entirely clear. Here we investigate
the role of W6.48 in the neuropeptide Y1 receptor (Y1R).
Via NMR experiment and molecular dynamics simulations, we find that
on the one hand, W6.48 exhibits multiple rotameric conformations,
where simulations indicate that these are coupled to backbone structure.
On the other hand, mutation of W6.48 to alanine does not
prevent G-protein signaling, and agonist vs antagonist bound Y1R exhibits
the same W6.48 rotameric state, calling into question whether
its core function is to regulate Y1R activation. Further investigation
indicates that the W6.48 rotameric state restricts microstates
accessible by Y1R and impacts backbone dynamics. Using principal component
analysis of multiple MD trajectories, we determine the structural
similarity between Y1R for the various apo, NPY, NPY/Gi, and antagonist-bound
conformational states. We propose a role for W6.48 in regulating
Y1R binding, in which rotameric changes for W6.48 influence
ligand binding by favoring backbone structures in apo Y1R that are
similar to those observed for the active protein. Mutation of W6.48 then does not eliminate these structures but reduces their
prevalence. Therefore, W6.48 provides “fine-tuning”
of Y1R signaling; mutation of the 6.48 position leaves Y1R active
but “detuned” such that signaling is still possible,
but proper functioning is inhibited by a decrease in ligand binding
rate.

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