+データを開く
-基本情報
登録情報 | データベース: PDB / ID: 8gvx | ||||||
---|---|---|---|---|---|---|---|
タイトル | Cryo-EM structure of the human TRPC5 ion channel in complex with G alpha i3 subunits, class2 | ||||||
要素 |
| ||||||
キーワード | METAL TRANSPORT / TRP / transient receptor potential | ||||||
機能・相同性 | 機能・相同性情報 regulation of membrane hyperpolarization / phosphatidylserine exposure on apoptotic cell surface / Role of second messengers in netrin-1 signaling / negative regulation of dendrite morphogenesis / store-operated calcium channel activity / cation channel complex / inositol 1,4,5 trisphosphate binding / negative regulation of adenylate cyclase activity / actinin binding / GTP metabolic process ...regulation of membrane hyperpolarization / phosphatidylserine exposure on apoptotic cell surface / Role of second messengers in netrin-1 signaling / negative regulation of dendrite morphogenesis / store-operated calcium channel activity / cation channel complex / inositol 1,4,5 trisphosphate binding / negative regulation of adenylate cyclase activity / actinin binding / GTP metabolic process / TRP channels / G protein-coupled dopamine receptor signaling pathway / clathrin binding / positive regulation of macroautophagy / Adenylate cyclase inhibitory pathway / positive regulation of axon extension / regulation of cytosolic calcium ion concentration / calcium channel complex / positive regulation of neuron differentiation / adenylate cyclase-inhibiting G protein-coupled receptor signaling pathway / G-protein beta/gamma-subunit complex binding / positive regulation of peptidyl-threonine phosphorylation / G protein-coupled receptor binding / calcium ion transmembrane transport / calcium channel activity / neuron differentiation / adenylate cyclase-modulating G protein-coupled receptor signaling pathway / ADP signalling through P2Y purinoceptor 12 / G alpha (z) signalling events / ADORA2B mediated anti-inflammatory cytokines production / GPER1 signaling / calcium ion transport / GDP binding / heterotrimeric G-protein complex / nervous system development / actin binding / positive regulation of cytosolic calcium ion concentration / ATPase binding / G alpha (i) signalling events / midbody / growth cone / G alpha (s) signalling events / neuron apoptotic process / Extra-nuclear estrogen signaling / lysosomal membrane / cell division / GTPase activity / centrosome / neuronal cell body / dendrite / positive regulation of cell population proliferation / GTP binding / nucleolus / Golgi apparatus / extracellular exosome / nucleoplasm / membrane / metal ion binding / plasma membrane / cytoplasm 類似検索 - 分子機能 | ||||||
生物種 | Homo sapiens (ヒト) | ||||||
手法 | 電子顕微鏡法 / 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 3.91 Å | ||||||
データ登録者 | Won, J. / Jeong, H. / Lee, H.H. | ||||||
資金援助 | 韓国, 1件
| ||||||
引用 | ジャーナル: Nat Commun / 年: 2023 タイトル: Molecular architecture of the Gα-bound TRPC5 ion channel. 著者: Jongdae Won / Jinsung Kim / Hyeongseop Jeong / Jinhyeong Kim / Shasha Feng / Byeongseok Jeong / Misun Kwak / Juyeon Ko / Wonpil Im / Insuk So / Hyung Ho Lee / 要旨: G-protein coupled receptors (GPCRs) and ion channels serve as key molecular switches through which extracellular stimuli are transformed into intracellular effects, and it has long been postulated ...G-protein coupled receptors (GPCRs) and ion channels serve as key molecular switches through which extracellular stimuli are transformed into intracellular effects, and it has long been postulated that ion channels are direct effector molecules of the alpha subunit of G-proteins (Gα). However, no complete structural evidence supporting the direct interaction between Gα and ion channels is available. Here, we present the cryo-electron microscopy structures of the human transient receptor potential canonical 5 (TRPC5)-Gα complexes with a 4:4 stoichiometry in lipid nanodiscs. Remarkably, Gα binds to the ankyrin repeat edge of TRPC5 ~ 50 Å away from the cell membrane. Electrophysiological analysis shows that Gα increases the sensitivity of TRPC5 to phosphatidylinositol 4,5-bisphosphate (PIP), thereby rendering TRPC5 more easily opened in the cell membrane, where the concentration of PIP is physiologically regulated. Our results demonstrate that ion channels are one of the direct effector molecules of Gα proteins triggered by GPCR activation-providing a structural framework for unraveling the crosstalk between two major classes of transmembrane proteins: GPCRs and ion channels. | ||||||
履歴 |
|
-構造の表示
構造ビューア | 分子: MolmilJmol/JSmol |
---|
-ダウンロードとリンク
-ダウンロード
PDBx/mmCIF形式 | 8gvx.cif.gz | 725.8 KB | 表示 | PDBx/mmCIF形式 |
---|---|---|---|---|
PDB形式 | pdb8gvx.ent.gz | 606.2 KB | 表示 | PDB形式 |
PDBx/mmJSON形式 | 8gvx.json.gz | ツリー表示 | PDBx/mmJSON形式 | |
その他 | その他のダウンロード |
-検証レポート
文書・要旨 | 8gvx_validation.pdf.gz | 1.5 MB | 表示 | wwPDB検証レポート |
---|---|---|---|---|
文書・詳細版 | 8gvx_full_validation.pdf.gz | 1.6 MB | 表示 | |
XML形式データ | 8gvx_validation.xml.gz | 109.2 KB | 表示 | |
CIF形式データ | 8gvx_validation.cif.gz | 157.1 KB | 表示 | |
アーカイブディレクトリ | https://data.pdbj.org/pub/pdb/validation_reports/gv/8gvx ftp://data.pdbj.org/pub/pdb/validation_reports/gv/8gvx | HTTPS FTP |
-関連構造データ
-リンク
-集合体
登録構造単位 |
|
---|---|
1 |
|
-要素
-タンパク質 , 2種, 8分子 BACDFEGH
#1: タンパク質 | 分子量: 89951.891 Da / 分子数: 4 / 由来タイプ: 組換発現 詳細: residues 766,767(SR) restriction enzyme, XbaI, residues 768-773(LEVLFQ) protease cleavage site, HRV-3C 由来: (組換発現) Homo sapiens (ヒト) / 遺伝子: TRPC5, TRP5 / 細胞株 (発現宿主): HEK293 / 発現宿主: Homo sapiens (ヒト) / 参照: UniProt: Q9UL62 #2: タンパク質 | 分子量: 41399.047 Da / 分子数: 4 / Mutation: Q204L / 由来タイプ: 組換発現 / 詳細: 6 histidine tag is inserted between M119 and T120. / 由来: (組換発現) Homo sapiens (ヒト) / 遺伝子: GNAI3 / 発現宿主: Escherichia coli K-12 (大腸菌) / 株 (発現宿主): K-12 / 参照: UniProt: P08754 |
---|
-非ポリマー , 6種, 24分子
#3: 化合物 | ChemComp-PTY / #4: 化合物 | ChemComp-Y01 / #5: 化合物 | ChemComp-ZN / #6: 化合物 | ChemComp-CA / #7: 化合物 | ChemComp-YZY / ( #8: 化合物 | ChemComp-GTP / |
---|
-詳細
研究の焦点であるリガンドがあるか | Y |
---|
-実験情報
-実験
実験 | 手法: 電子顕微鏡法 |
---|---|
EM実験 | 試料の集合状態: PARTICLE / 3次元再構成法: 単粒子再構成法 |
-試料調製
構成要素 | 名称: Transient receptor potential / タイプ: COMPLEX / Entity ID: #1 / 由来: RECOMBINANT |
---|---|
分子量 | 実験値: NO |
由来(天然) | 生物種: Homo sapiens (ヒト) |
由来(組換発現) | 生物種: Homo sapiens (ヒト) / 細胞: HEK293 |
緩衝液 | pH: 8 |
試料 | 包埋: NO / シャドウイング: NO / 染色: NO / 凍結: YES |
急速凍結 | 凍結剤: ETHANE |
-電子顕微鏡撮影
顕微鏡 | モデル: TFS GLACIOS |
---|---|
電子銃 | 電子線源: FIELD EMISSION GUN / 加速電圧: 200 kV / 照射モード: FLOOD BEAM |
電子レンズ | モード: BRIGHT FIELD / 最大 デフォーカス(公称値): 2000 nm / 最小 デフォーカス(公称値): 1000 nm |
撮影 | 電子線照射量: 40 e/Å2 フィルム・検出器のモデル: FEI FALCON IV (4k x 4k) |
-解析
ソフトウェア | 名称: PHENIX / バージョン: 1.19.2_4158: / 分類: 精密化 | ||||||||||||||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
CTF補正 | タイプ: PHASE FLIPPING AND AMPLITUDE CORRECTION | ||||||||||||||||||||||||
3次元再構成 | 解像度: 3.91 Å / 解像度の算出法: OTHER / 粒子像の数: 5344 詳細: We combined two different maps from the same dataset (D_1300031718_em-additional-volume_P1.map.V4 and D_1300031718_em-additional-volume_P2.map.V4) to generate a composite map (D_1300031718_em- ...詳細: We combined two different maps from the same dataset (D_1300031718_em-additional-volume_P1.map.V4 and D_1300031718_em-additional-volume_P2.map.V4) to generate a composite map (D_1300031718_em-volume_P1.map.V6). The density of the G protein area could not be visualized clearly in the consensus map of this EM dataset. Therefore, we performed focused classification and local refinement to improve the density of the G protein area using symmetry expanded particles with C4 symmetry imposition, which required more number of particles. Finally, the number of particles used to reconstruct additional volume data 1 (D_1300031718_em-additional-volume_P1.map.V4) is 5,344 and the number of particles used to reconstruct additional volume data 2 (D_1300031718_em-additional-volume_P2.map.V4) is 205,343. Furthermore, the resolution stated above is based on map resolution estimates calculated by a validation tool in Phenix, FSC (model) = 0.143. 対称性のタイプ: POINT | ||||||||||||||||||||||||
拘束条件 |
|