The transient receptor potential vanilloid 2 (TRPV2) ion channel is a polymodal receptor widely involved in many physiological and pathological processes. Despite many TRPV2 modulators being identified, whether and how TRPV2 is regulated by endogenous lipids remains elusive. Here, we report an endogenous cholesterol molecule inside the vanilloid binding pocket (VBP) of TRPV2, with a ‘head down, tail up’ configuration, resolved at 3.2 Å using cryo-EM. Cholesterol binding antagonizes ligand activation of TRPV2, which is removed from VBP by methyl-β-cyclodextrin (MβCD) as resolved at 2.9 Å. We also observed that estradiol (E2) potentiated TRPV2 activation by 2-aminoethoxydiphenyl borate (2-APB), a classic tool compound for TRP channels. Our cryo-EM structures (resolved at 2.8–3.3 Å) further suggest how E2 disturbed cholesterol binding and how 2-APB bound within the VBP with E2 or without both E2 and endogenous cholesterol, respectively. Therefore, our study has established the structural basis for ligand recognition of the inhibitory endogenous cholesterol and excitatory exogenous 2-APB in TRPV2.

瞬时受体电位香草素 2 (TRPV2) 离子通道是一种多模式受体，广泛参与许多生理和病理过程。尽管鉴定了许多 TRPV2 调节剂，但 TRPV2 是否以及如何受内源性脂质调节仍然难以捉摸。在这里，我们报告了 TRPV2 的香草素结合袋 (VBP) 内的内源性胆固醇分子，具有“头朝下，尾巴朝上”的配置，使用冷冻电子显微镜在 3.2 Å 处分辨。胆固醇结合拮抗 TRPV2 的配体激活，TRPV2 通过甲基-β-环糊精 (MβCD) 从 VBP 中去除，分辨率为 2.9 Å。我们还观察到雌二醇 (E2) 通过 2-氨基乙氧基二苯基硼酸酯 (2-APB) 增强 TRPV2 激活，2-APB 是 TRP 通道的经典工具化合物。我们的冷冻电镜结构（分辨率为 2.8–3. 3 Å) 进一步表明 E2 如何干扰胆固醇结合以及 2-APB 如何在 VBP 内结合 E2 或不结合 E2 和内源性胆固醇，分别。因此，我们的研究建立了配体识别 TRPV2 中抑制性内源性胆固醇和兴奋性外源性 2-APB 的结构基础。