Transient receptor potential (TRP) channel TRPV4 is a polymodal cellular sensor that responds to moderate heat, cell swelling, shear stress, and small-molecule ligands. It is involved in thermogenesis, regulation of vascular tone, bone homeostasis, renal and pulmonary functions. TRPV4 is implicated in neuromuscular and skeletal disorders, pulmonary edema, and cancers, and represents an important drug target. The cytoskeletal remodeling GTPase RhoA has been shown to suppress TRPV4 activity. Here, we present a structure of the human TRPV4-RhoA complex that shows RhoA interaction with the membrane-facing surface of the TRPV4 ankyrin repeat domains. The contact interface reveals residues that are mutated in neuropathies, providing an insight into the disease pathogenesis. We also identify the binding sites of the TRPV4 agonist 4α-PDD and the inhibitor HC-067047 at the base of the S1-S4 bundle, and show that agonist binding leads to pore opening, while channel inhibition involves a π-to-α transition in the pore-forming helix S6. Our structures elucidate the interaction interface between hTRPV4 and RhoA, as well as residues at this interface that are involved in TRPV4 disease-causing mutations. They shed light on TRPV4 activation and inhibition and provide a template for the design of future therapeutics for treatment of TRPV4-related diseases.

瞬时受体电位 (TRP) 通道 TRPV4 是一种多模式细胞传感器，可响应适度的热量、细胞肿胀、剪切应力和小分子配体。它参与产热、血管张力调节、骨稳态、肾功能和肺功能。TRPV4 与神经肌肉和骨骼疾病、肺水肿和癌症有关，是一个重要的药物靶点。细胞骨架重塑 GTPase RhoA 已被证明可以抑制 TRPV4 活性。在这里，我们展示了人类 TRPV4-RhoA 复合物的结构，该结构显示了 RhoA 与 TRPV4 锚蛋白重复结构域的面向膜表面的相互作用。接触界面揭示了神经病中突变的残基，提供了对疾病发病机制的深入了解。我们还确定了 TRPV4 激动剂 4α-PDD 和抑制剂 HC-067047 在 S1-S4 束底部的结合位点，并表明激动剂结合导致孔打开，而通道抑制涉及 π 到 α 的转变在成孔螺旋 S6 中。我们的结构阐明了 hTRPV4 和 RhoA 之间的相互作用界面，以及该界面上与 TRPV4 致病突变有关的残基。它们揭示了 TRPV4 的激活和抑制，并为未来治疗 TRPV4 相关疾病的疗法的设计提供了模板。