Japan Agency for Medical Research and Development (AMED)
JP20ae0101051
日本
Ministry of Education, Culture, Sports, Science and Technology (Japan)
JP20H03165
日本
New Energy and Industrial Technology Development Organization (NEDO)
JP20am0101118
日本
引用
ジャーナル: Sci Signal / 年: 2022 タイトル: Structures of human pannexin-1 in nanodiscs reveal gating mediated by dynamic movement of the N terminus and phospholipids. 著者: Maki Kuzuya / Hidemi Hirano / Kenichi Hayashida / Masakatsu Watanabe / Kazumi Kobayashi / Tohru Terada / Md Iqbal Mahmood / Florence Tama / Kazutoshi Tani / Yoshinori Fujiyoshi / Atsunori Oshima / 要旨: Pannexin (PANX) family proteins form large-pore channels that mediate purinergic signaling. We analyzed the cryo-EM structures of human PANX1 in lipid nanodiscs to elucidate the gating mechanism and ...Pannexin (PANX) family proteins form large-pore channels that mediate purinergic signaling. We analyzed the cryo-EM structures of human PANX1 in lipid nanodiscs to elucidate the gating mechanism and its regulation by the amino terminus in phospholipids. The wild-type channel has an amino-terminal funnel in the pore, but in the presence of the inhibitor probenecid, a cytoplasmically oriented amino terminus and phospholipids obstruct the pore. Functional analysis using whole-cell patch-clamp and oocyte voltage clamp showed that PANX1 lacking the amino terminus did not open and had a dominant negative effect on channel activity, thus confirming that the amino-terminal domain played an essential role in channel opening. These observations suggest that dynamic conformational changes in the amino terminus of human PANX1 are associated with lipid movement in and out of the pore. Moreover, the data provide insight into the gating mechanism of PANX1 and, more broadly, other large-pore channels.
EMPIAR-10759 (タイトル: Structure of C-terminal deleted human pannexin-1 in nanodisc Data size: 784.4 Data #1: C-terminal deletion mutant of human PANX1 in nanodisc [micrographs - multiframe])
凍結剤: ETHANE / 装置: LEICA KF80 詳細: blot for 10 seconds at room temperature followed by plunge freezing. Because of manual blotting, humidity and temperature in a room is not controlled..