+データを開く
-基本情報
登録情報 | データベース: PDB / ID: 8t1e | |||||||||||||||||||||
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タイトル | Closed-state cryo-EM structure of full-length human TRPV4 in the presence of 4a-PDD | |||||||||||||||||||||
要素 | Transient receptor potential cation channel subfamily V member 4,Enhanced green fluorescent protein | |||||||||||||||||||||
キーワード | MEMBRANE PROTEIN (膜タンパク質) / transient receptor potential V family member 4 / TRP / channel / TRPV4 / TRP channels (TRPチャネル) / agonist (アゴニスト) / 4a-PDD / 4alpha-Phorbol 12 / 13-didecanoate / closed state | |||||||||||||||||||||
機能・相同性 | 機能・相同性情報 stretch-activated, monoatomic cation-selective, calcium channel activity / blood vessel endothelial cell delamination / osmosensor activity / vasopressin secretion / positive regulation of striated muscle contraction / calcium ion import into cytosol / positive regulation of macrophage inflammatory protein 1 alpha production / negative regulation of brown fat cell differentiation / positive regulation of microtubule depolymerization / hyperosmotic salinity response ...stretch-activated, monoatomic cation-selective, calcium channel activity / blood vessel endothelial cell delamination / osmosensor activity / vasopressin secretion / positive regulation of striated muscle contraction / calcium ion import into cytosol / positive regulation of macrophage inflammatory protein 1 alpha production / negative regulation of brown fat cell differentiation / positive regulation of microtubule depolymerization / hyperosmotic salinity response / cortical microtubule organization / positive regulation of chemokine (C-X-C motif) ligand 1 production / positive regulation of chemokine (C-C motif) ligand 5 production / cartilage development involved in endochondral bone morphogenesis / cellular hypotonic response / regulation of response to osmotic stress / cellular hypotonic salinity response / osmosensory signaling pathway / multicellular organismal-level water homeostasis / positive regulation of vascular permeability / cellular response to osmotic stress / calcium ion import / cell volume homeostasis / positive regulation of monocyte chemotactic protein-1 production / cell-cell junction assembly / TRPチャネル / regulation of aerobic respiration / cortical actin cytoskeleton / positive regulation of macrophage chemotaxis / beta-tubulin binding / diet induced thermogenesis / 微小管 / alpha-tubulin binding / cytoplasmic microtubule / response to mechanical stimulus / monoatomic cation channel activity / SH2 domain binding / 生物発光 / protein kinase C binding / filopodium / generation of precursor metabolites and energy / actin filament organization / calcium ion transmembrane transport / 接着結合 / positive regulation of JNK cascade / response to insulin / calcium channel activity / 繊毛 / ruffle membrane / intracellular calcium ion homeostasis / positive regulation of inflammatory response / positive regulation of interleukin-6 production / calcium ion transport / actin filament binding / glucose homeostasis / negative regulation of neuron projection development / lamellipodium / cellular response to heat / actin binding / 成長円錐 / positive regulation of cytosolic calcium ion concentration / actin cytoskeleton organization / microtubule binding / response to hypoxia / positive regulation of ERK1 and ERK2 cascade / calmodulin binding / apical plasma membrane / focal adhesion / lipid binding / protein kinase binding / negative regulation of transcription by RNA polymerase II / 細胞膜 / 小胞体 / ATP binding / 生体膜 / identical protein binding / metal ion binding / 細胞膜 類似検索 - 分子機能 | |||||||||||||||||||||
生物種 | Homo sapiens (ヒト) Human betaherpesvirus 5 (ヘルペスウイルス) | |||||||||||||||||||||
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: 2.77 Å | |||||||||||||||||||||
データ登録者 | Talyzina, I.A. / Nadezhdin, K.D. / Neuberger, A. / Sobolevsky, A.I. | |||||||||||||||||||||
資金援助 | 米国, ドイツ, 6件
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引用 | ジャーナル: Nat Commun / 年: 2023 タイトル: Structure of human TRPV4 in complex with GTPase RhoA. 著者: Kirill D Nadezhdin / Irina A Talyzina / Aravind Parthasarathy / Arthur Neuberger / David X Zhang / Alexander I Sobolevsky / 要旨: 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 ...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. | |||||||||||||||||||||
履歴 |
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-構造の表示
構造ビューア | 分子: MolmilJmol/JSmol |
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-ダウンロードとリンク
-ダウンロード
PDBx/mmCIF形式 | 8t1e.cif.gz | 513.6 KB | 表示 | PDBx/mmCIF形式 |
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PDB形式 | pdb8t1e.ent.gz | 412.7 KB | 表示 | PDB形式 |
PDBx/mmJSON形式 | 8t1e.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
アーカイブディレクトリ | https://data.pdbj.org/pub/pdb/validation_reports/t1/8t1e ftp://data.pdbj.org/pub/pdb/validation_reports/t1/8t1e | HTTPS FTP |
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-関連構造データ
関連構造データ | 40961MC 8t1bC 8t1cC 8t1dC 8t1fC C: 同じ文献を引用 (文献) M: このデータのモデリングに利用したマップデータ |
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類似構造データ | 類似検索 - 機能・相同性F&H 検索 |
-リンク
-集合体
登録構造単位 |
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1 |
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-要素
#1: タンパク質 | 分子量: 127717.938 Da / 分子数: 4 / 由来タイプ: 組換発現 由来: (組換発現) Homo sapiens (ヒト), (組換発現) Human betaherpesvirus 5 (ヘルペスウイルス) 遺伝子: TRPV4, VRL2, VROAC, egfp / 細胞株 (発現宿主): HEK293S / 発現宿主: Homo sapiens (ヒト) / 参照: UniProt: Q9HBA0, UniProt: C5MKY7 #2: 化合物 | ChemComp-9ZR / [( #3: 化合物 | ChemComp-YJ0 / ( #4: 化合物 | #5: 水 | ChemComp-HOH / | 研究の焦点であるリガンドがあるか | N | |
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-実験情報
-実験
実験 | 手法: 電子顕微鏡法 |
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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: full-length human TRPV4 in the presence of 4a-PDD / タイプ: COMPLEX / Entity ID: #1 / 由来: RECOMBINANT | ||||||||||||||||||||||||||||||
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分子量 | 値: 0.51 MDa / 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: Human embryonic kidney 293 / プラスミド: pEG BacMam | ||||||||||||||||||||||||||||||
緩衝液 | pH: 8 | ||||||||||||||||||||||||||||||
緩衝液成分 |
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試料 | 濃度: 3.2 mg/ml / 包埋: NO / シャドウイング: NO / 染色: NO / 凍結: YES / 詳細: human TRPV4 | ||||||||||||||||||||||||||||||
試料支持 | グリッドの材料: GOLD / グリッドのサイズ: 300 divisions/in. / グリッドのタイプ: UltrAuFoil R1.2/1.3 | ||||||||||||||||||||||||||||||
急速凍結 | 装置: FEI VITROBOT MARK IV / 凍結剤: ETHANE / 湿度: 100 % / 凍結前の試料温度: 295 K |
-電子顕微鏡撮影
実験機器 | モデル: Titan Krios / 画像提供: FEI Company |
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顕微鏡 | モデル: TFS KRIOS |
電子銃 | 電子線源: FIELD EMISSION GUN / 加速電圧: 300 kV / 照射モード: FLOOD BEAM |
電子レンズ | モード: BRIGHT FIELDBright-field microscopy / 最大 デフォーカス(公称値): 1500 nm / 最小 デフォーカス(公称値): 750 nm / Cs: 2.7 mm |
撮影 | 平均露光時間: 2.4 sec. / 電子線照射量: 60 e/Å2 / フィルム・検出器のモデル: GATAN K3 (6k x 4k) / 撮影したグリッド数: 1 / 実像数: 15624 |
画像スキャン | 横: 11520 / 縦: 8184 |
-解析
EMソフトウェア | 名称: PHENIX / バージョン: 1.11.1_2575: / カテゴリ: モデル精密化 | ||||||||||||||||||||||||
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CTF補正 | タイプ: NONE | ||||||||||||||||||||||||
粒子像の選択 | 選択した粒子像数: 3935964 | ||||||||||||||||||||||||
3次元再構成 | 解像度: 2.77 Å / 解像度の算出法: FSC 0.143 CUT-OFF / 粒子像の数: 75146 / 対称性のタイプ: POINT | ||||||||||||||||||||||||
原子モデル構築 | 空間: REAL | ||||||||||||||||||||||||
拘束条件 |
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