+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-25492 | |||||||||
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タイトル | Cryo-EM structure of the human NALCN channelosome NALCN-FAM155A-CaM-UNC79-UNC80 complex conformation 1 | |||||||||
マップデータ | combined map of focused refinements used for model refinements | |||||||||
試料 |
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キーワード | ion channel / calmodulin / HEAT repeat protein / MEMBRANE PROTEIN | |||||||||
機能・相同性 | 機能・相同性情報 monoatomic cation homeostasis / positive regulation of synaptic transmission, cholinergic / leak channel activity / regulation of resting membrane potential / cation channel complex / CaM pathway / Cam-PDE 1 activation / voltage-gated sodium channel activity / Sodium/Calcium exchangers / sodium channel activity ...monoatomic cation homeostasis / positive regulation of synaptic transmission, cholinergic / leak channel activity / regulation of resting membrane potential / cation channel complex / CaM pathway / Cam-PDE 1 activation / voltage-gated sodium channel activity / Sodium/Calcium exchangers / sodium channel activity / Calmodulin induced events / Reduction of cytosolic Ca++ levels / CREB1 phosphorylation through the activation of CaMKII/CaMKK/CaMKIV cascasde / Activation of Ca-permeable Kainate Receptor / Loss of phosphorylation of MECP2 at T308 / CREB1 phosphorylation through the activation of Adenylate Cyclase / PKA activation / negative regulation of high voltage-gated calcium channel activity / monoatomic ion channel complex / CaMK IV-mediated phosphorylation of CREB / Glycogen breakdown (glycogenolysis) / positive regulation of cyclic-nucleotide phosphodiesterase activity / organelle localization by membrane tethering / negative regulation of calcium ion export across plasma membrane / CLEC7A (Dectin-1) induces NFAT activation / autophagosome membrane docking / mitochondrion-endoplasmic reticulum membrane tethering / Activation of RAC1 downstream of NMDARs / regulation of cardiac muscle cell action potential / positive regulation of ryanodine-sensitive calcium-release channel activity / calcium ion import across plasma membrane / regulation of cell communication by electrical coupling involved in cardiac conduction / Synthesis of IP3 and IP4 in the cytosol / negative regulation of peptidyl-threonine phosphorylation / Negative regulation of NMDA receptor-mediated neuronal transmission / Phase 0 - rapid depolarisation / Unblocking of NMDA receptors, glutamate binding and activation / negative regulation of ryanodine-sensitive calcium-release channel activity / protein phosphatase activator activity / RHO GTPases activate PAKs / Ion transport by P-type ATPases / : / Uptake and function of anthrax toxins / Long-term potentiation / Regulation of MECP2 expression and activity / Calcineurin activates NFAT / catalytic complex / DARPP-32 events / detection of calcium ion / regulation of cardiac muscle contraction / Smooth Muscle Contraction / regulation of ryanodine-sensitive calcium-release channel activity / RHO GTPases activate IQGAPs / regulation of cardiac muscle contraction by regulation of the release of sequestered calcium ion / sodium ion transmembrane transport / calcium channel inhibitor activity / cellular response to interferon-beta / eNOS activation / Protein methylation / monoatomic cation channel activity / voltage-gated potassium channel complex / Activation of AMPK downstream of NMDARs / regulation of release of sequestered calcium ion into cytosol by sarcoplasmic reticulum / Tetrahydrobiopterin (BH4) synthesis, recycling, salvage and regulation / Ion homeostasis / : / titin binding / positive regulation of protein autophosphorylation / regulation of calcium-mediated signaling / sperm midpiece / calcium channel complex / potassium ion transmembrane transport / substantia nigra development / adenylate cyclase activator activity / Ras activation upon Ca2+ influx through NMDA receptor / regulation of heart rate / sarcomere / FCERI mediated Ca+2 mobilization / FCGR3A-mediated IL10 synthesis / protein serine/threonine kinase activator activity / bioluminescence / VEGFR2 mediated vascular permeability / VEGFR2 mediated cell proliferation / regulation of cytokinesis / Antigen activates B Cell Receptor (BCR) leading to generation of second messengers / positive regulation of peptidyl-threonine phosphorylation / generation of precursor metabolites and energy / spindle microtubule / positive regulation of synaptic transmission, GABAergic / Translocation of SLC2A4 (GLUT4) to the plasma membrane / positive regulation of receptor signaling pathway via JAK-STAT / calcium ion transmembrane transport / RAF activation / Transcriptional activation of mitochondrial biogenesis / positive regulation of protein serine/threonine kinase activity / Stimuli-sensing channels / cellular response to type II interferon / spindle pole / response to calcium ion / RAS processing 類似検索 - 分子機能 | |||||||||
生物種 | Homo sapiens (ヒト) | |||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 3.1 Å | |||||||||
データ登録者 | Kschonsak M / Chua HC | |||||||||
資金援助 | 1件
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引用 | ジャーナル: Nature / 年: 2022 タイトル: Structural architecture of the human NALCN channelosome. 著者: Marc Kschonsak / Han Chow Chua / Claudia Weidling / Nourdine Chakouri / Cameron L Noland / Katharina Schott / Timothy Chang / Christine Tam / Nidhi Patel / Christopher P Arthur / Alexander ...著者: Marc Kschonsak / Han Chow Chua / Claudia Weidling / Nourdine Chakouri / Cameron L Noland / Katharina Schott / Timothy Chang / Christine Tam / Nidhi Patel / Christopher P Arthur / Alexander Leitner / Manu Ben-Johny / Claudio Ciferri / Stephan Alexander Pless / Jian Payandeh / 要旨: Depolarizing sodium (Na) leak currents carried by the NALCN channel regulate the resting membrane potential of many neurons to modulate respiration, circadian rhythm, locomotion and pain sensitivity. ...Depolarizing sodium (Na) leak currents carried by the NALCN channel regulate the resting membrane potential of many neurons to modulate respiration, circadian rhythm, locomotion and pain sensitivity. NALCN requires FAM155A, UNC79 and UNC80 to function, but the role of these auxiliary subunits is not understood. NALCN, UNC79 and UNC80 are essential in rodents, and mutations in human NALCN and UNC80 cause severe developmental and neurological disease. Here we determined the structure of the NALCN channelosome, an approximately 1-MDa complex, as fundamental aspects about the composition, assembly and gating of this channelosome remain obscure. UNC79 and UNC80 are massive HEAT-repeat proteins that form an intertwined anti-parallel superhelical assembly, which docks intracellularly onto the NALCN-FAM155A pore-forming subcomplex. Calmodulin copurifies bound to the carboxy-terminal domain of NALCN, identifying this region as a putative modulatory hub. Single-channel analyses uncovered a low open probability for the wild-type complex, highlighting the tightly closed S6 gate in the structure, and providing a basis to interpret the altered gating properties of disease-causing variants. Key constraints between the UNC79-UNC80 subcomplex and the NALCN DI-DII and DII-DIII linkers were identified, leading to a model of channelosome gating. Our results provide a structural blueprint to understand the physiology of the NALCN channelosome and a template for drug discovery to modulate the resting membrane potential. | |||||||||
履歴 |
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-構造の表示
ムービー |
ムービービューア |
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構造ビューア | EMマップ: SurfViewMolmilJmol/JSmol |
添付画像 |
-ダウンロードとリンク
-EMDBアーカイブ
マップデータ | emd_25492.map.gz | 221.2 MB | EMDBマップデータ形式 | |
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ヘッダ (付随情報) | emd-25492-v30.xml emd-25492.xml | 68.6 KB 68.6 KB | 表示 表示 | EMDBヘッダ |
画像 | emd_25492.png | 90.3 KB | ||
Filedesc metadata | emd-25492.cif.gz | 13.3 KB | ||
その他 | emd_25492_additional_1.map.gz emd_25492_additional_10.map.gz emd_25492_additional_11.map.gz emd_25492_additional_12.map.gz emd_25492_additional_13.map.gz emd_25492_additional_14.map.gz emd_25492_additional_15.map.gz emd_25492_additional_16.map.gz emd_25492_additional_17.map.gz emd_25492_additional_2.map.gz emd_25492_additional_3.map.gz emd_25492_additional_4.map.gz emd_25492_additional_5.map.gz emd_25492_additional_6.map.gz emd_25492_additional_7.map.gz emd_25492_additional_8.map.gz emd_25492_additional_9.map.gz emd_25492_half_map_1.map.gz emd_25492_half_map_2.map.gz | 226.1 MB 89.6 MB 89.6 MB 226.9 MB 225.9 MB 89.6 MB 89.6 MB 226.9 MB 89.6 MB 89.6 MB 226.1 MB 226.8 MB 226 MB 89.6 MB 89.6 MB 226.5 MB 226.2 MB 89.6 MB 89.6 MB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-25492 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-25492 | HTTPS FTP |
-検証レポート
文書・要旨 | emd_25492_validation.pdf.gz | 1021.7 KB | 表示 | EMDB検証レポート |
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文書・詳細版 | emd_25492_full_validation.pdf.gz | 1021.2 KB | 表示 | |
XML形式データ | emd_25492_validation.xml.gz | 16.2 KB | 表示 | |
CIF形式データ | emd_25492_validation.cif.gz | 19.4 KB | 表示 | |
アーカイブディレクトリ | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-25492 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-25492 | HTTPS FTP |
-関連構造データ
-リンク
EMDBのページ | EMDB (EBI/PDBe) / EMDataResource |
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「今月の分子」の関連する項目 |
-マップ
ファイル | ダウンロード / ファイル: emd_25492.map.gz / 形式: CCP4 / 大きさ: 244.1 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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注釈 | combined map of focused refinements used for model refinements | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
投影像・断面図 | 画像のコントロール
画像は Spider により作成 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 1.1732 Å | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
密度 |
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対称性 | 空間群: 1 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
詳細 | EMDB XML:
CCP4マップ ヘッダ情報:
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-添付データ
+追加マップ: map (non-sharpened) of focused NALCN-FAM region containing the...
+追加マップ: halfmap 2 of focused UNC-crossover region containing the...
+追加マップ: halfmap 1 of focused UNC-crossover region containing the...
+追加マップ: sharpened map of focused UNC-CN region containing the...
+追加マップ: map (non-sharpened) of focused UNC-CN region containing the...
+追加マップ: halfmap 2 of focused UNC-CN region containing the...
+追加マップ: halfmap 1 of focused UNC-CN region containing the...
+追加マップ: sharpened map of focused NALCN-FAM region containing the...
+追加マップ: halfmap 1 of focused NALCN-FAM region containing the...
+追加マップ: halfmap 2 of focused NALCN-FAM region containing the...
+追加マップ: map (non-sharpened) of overall NALCN-FAM-UNC79-UNC80-CAM used for alignment...
+追加マップ: sharpened map of focused UNC-NC region containing the...
+追加マップ: map (non-sharpened) of focused UNC-NC region containing the...
+追加マップ: halfmap 2 of focused UNC-NC region containing the...
+追加マップ: halfmap 1 of focused UNC-NC region containing the...
+追加マップ: sharpened map of focused UNC-crossover region containing the...
+追加マップ: map (non-sharpened) of focused UNC-crossover region containing the...
+ハーフマップ: halfmap 1 (non-sharpened) of overall NALCN-FAM-UNC79-UNC80-CAM
+ハーフマップ: halfmap 2 (non-sharpened) of overall NALCN-FAM-UNC79-UNC80-CAM
-試料の構成要素
+全体 : Pentameric complex of the leak channel NALCN with FAM155A, Calmod...
+超分子 #1: Pentameric complex of the leak channel NALCN with FAM155A, Calmod...
+分子 #1: Sodium leak channel non-selective protein,Enhanced green fluoresc...
+分子 #2: Transmembrane protein FAM155A
+分子 #3: Calmodulin-1
+分子 #4: UNC79,Protein unc-79 homolog,Protein unc-79 homolog
+分子 #5: Protein unc-80 homolog
+分子 #6: 2-acetamido-2-deoxy-beta-D-glucopyranose
+分子 #7: (1S)-2-{[(2-AMINOETHOXY)(HYDROXY)PHOSPHORYL]OXY}-1-[(PALMITOYLOXY...
+分子 #8: (1R)-2-{[{[(2S)-2,3-DIHYDROXYPROPYL]OXY}(HYDROXY)PHOSPHORYL]OXY}-...
+分子 #9: CHOLESTEROL HEMISUCCINATE
-実験情報
-構造解析
手法 | クライオ電子顕微鏡法 |
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解析 | 単粒子再構成法 |
試料の集合状態 | particle |
-試料調製
濃度 | 1.65 mg/mL | |||||||||
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緩衝液 | pH: 7.5 構成要素:
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グリッド | モデル: UltrAuFoil R0.6/1 / 材質: GOLD / メッシュ: 300 / 前処理 - タイプ: PLASMA CLEANING / 前処理 - 時間: 30 sec. | |||||||||
凍結 | 凍結剤: ETHANE / チャンバー内湿度: 100 % / チャンバー内温度: 277 K / 装置: LEICA EM GP / 詳細: 3.5 sec blotting. | |||||||||
詳細 | NALCN was reconstituted into lipid nanodiscs and mildly crosslinkned with 0.05% EM grade glutaraldehyde for 10 min at room temperature. Cross-linking was quenched with 9 mM Tris pH 7.5 |
-電子顕微鏡法
顕微鏡 | FEI TITAN KRIOS |
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特殊光学系 | エネルギーフィルター - 名称: GIF Bioquantum / エネルギーフィルター - スリット幅: 20 eV |
撮影 | フィルム・検出器のモデル: GATAN K3 BIOQUANTUM (6k x 4k) 撮影したグリッド数: 2 / 実像数: 26550 / 平均露光時間: 3.0 sec. / 平均電子線量: 64.009 e/Å2 |
電子線 | 加速電圧: 300 kV / 電子線源: FIELD EMISSION GUN |
電子光学系 | 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD / Cs: 2.7 mm / 最大 デフォーカス(公称値): 1.5 µm / 最小 デフォーカス(公称値): 0.5 µm / 倍率(公称値): 105000 |
試料ステージ | 試料ホルダーモデル: FEI TITAN KRIOS AUTOGRID HOLDER ホルダー冷却材: NITROGEN |
実験機器 | モデル: Titan Krios / 画像提供: FEI Company |