Transient receptor potential (TRP) channels emerged in fungi as mechanosensitive osmoregulators. The Saccharomyces cerevisiae vacuolar TRP yeast 1 (TRPY1) is the most studied TRP channel from fungi, but the structure and details of channel modulation remain elusive. Here, we describe the full-length cryoelectron microscopy structure of TRPY1 at 3.1 Å resolution in a closed state. The structure, despite containing an evolutionarily conserved and archetypical transmembrane domain, reveals distinctive structural folds for the cytosolic N and C termini, compared with other eukaryotic TRP channels. We identify an inhibitory phosphatidylinositol 3-phosphate (PI(3)P) lipid-binding site, along with two Ca²⁺-binding sites: a cytosolic site, implicated in channel activation and a vacuolar lumen site, implicated in inhibition. These findings, together with data from microsecond-long molecular dynamics simulations and a model of a TRPY1 open state, provide insights into the basis of TRPY1 channel modulation by lipids and Ca²⁺, and the molecular evolution of TRP channels.

瞬时受体电位（TRP）通道在真菌中作为机械敏感渗透调节器出现。酿酒酵母液泡 TRP酵母1 (TRPY1) 是研究最多的真菌 TRP 通道，但通道调节的结构和细节仍然难以捉摸。在这里，我们以 3.1 Å 分辨率描述了关闭状态下 TRPY1 的全长冷冻电子显微镜结构。尽管该结构包含进化上保守的原型跨膜结构域，但与其他真核 TRP 通道相比，其胞浆 N 和 C 末端显示出独特的结构折叠。我们鉴定了一个抑制性磷脂酰肌醇 3-磷酸 (PI(3)P) 脂质结合位点，以及两个 Ca ²⁺结合位点：一个与通道激活有关的胞质位点和一个与抑制有关的液泡腔位点。^{这些发现与微秒长的分子动力学模拟数据和 TRPY1 开放状态模型一起，为了解脂质和 Ca 2+}调节 TRPY1 通道的基础以及 TRP 通道的分子进化提供了见解。