An Extracellular Pore‑Targeting Peptide Defines a Designable Allosteric Site in TRPV2

Extracellular pore domains of ion channels are emerging as dynamic regulatory surfaces, but whether they can be rationally targeted to achieve selective channel modulation remains unclear. Here, using an optimized hotspot-centric design strategy, we developed Depiv2, a structure-guided peptidic inhibitor of TRPV2 that binds the extracellular pore domain and suppresses channel activity with nanomolar potency and subtype selectivity. Our cryo-EM structure of the TRPV2-Depiv2 complex showed that peptide binding remodels the pore domain and stabilizes a closed, non-conductive conformation. Patch-clamp recordings further revealed that Depiv2 decreases both open probability and single-channel conductance, indicating inhibition through combined allosteric and permeation-coupled mechanisms. In cellular and mouse models of pathological cardiac hypertrophy, Depiv2 blunted disease-associated remodeling. These findings establish the extracellular pore of TRPV2 as a designable allosteric site and illustrate how rational peptide engineering can be used to target extracellular regulatory surfaces in TRP channels.