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基本情報
登録情報 | データベース: EMDB / ID: EMD-9025 | |||||||||
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タイトル | The voltage-activated Kv1.2-2.1 paddle chimera channel in lipid nanodiscs, cytosolic domain | |||||||||
![]() | Kv1.2-2.1 paddle chimera in lipid nanodiscs, cytoplasmic domain | |||||||||
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![]() | MEMBRANE PROTEIN / transport protein / potassium channel / lipid nanodisc | |||||||||
機能・相同性 | ![]() optic nerve structural organization / pinceau fiber / regulation of action potential / Voltage gated Potassium channels / voltage-gated monoatomic ion channel activity involved in regulation of postsynaptic membrane potential / potassium channel complex / NADPH oxidation / potassium ion export across plasma membrane / paranodal junction / regulation of circadian sleep/wake cycle, non-REM sleep ...optic nerve structural organization / pinceau fiber / regulation of action potential / Voltage gated Potassium channels / voltage-gated monoatomic ion channel activity involved in regulation of postsynaptic membrane potential / potassium channel complex / NADPH oxidation / potassium ion export across plasma membrane / paranodal junction / regulation of circadian sleep/wake cycle, non-REM sleep / axon initial segment / regulation of protein localization to cell surface / corpus callosum development / aldo-keto reductase (NADPH) activity / voltage-gated monoatomic ion channel activity involved in regulation of presynaptic membrane potential / juxtaparanode region of axon / regulation of potassium ion transmembrane transport / delayed rectifier potassium channel activity / outward rectifier potassium channel activity / 酸化還元酵素; CH-OHの結合に対し酸化酵素として働く; NAD又はNADPを用いる / myoblast differentiation / neuromuscular process / Neutrophil degranulation / optic nerve development / neuronal cell body membrane / regulation of dopamine secretion / voltage-gated potassium channel activity / kinesin binding / calyx of Held / lamellipodium membrane / potassium channel regulator activity / neuronal action potential / hematopoietic progenitor cell differentiation / axon terminus / potassium ion transmembrane transport / voltage-gated potassium channel complex / extrinsic component of cytoplasmic side of plasma membrane / sensory perception of pain / protein localization to plasma membrane / postsynaptic density membrane / potassium ion transport / protein homooligomerization / cytoplasmic side of plasma membrane / cerebral cortex development / lamellipodium / presynaptic membrane / postsynaptic membrane / perikaryon / transmembrane transporter binding / postsynaptic density / cytoskeleton / endosome / neuron projection / protein heterodimerization activity / axon / neuronal cell body / glutamatergic synapse / dendrite / protein-containing complex binding / endoplasmic reticulum membrane / membrane / plasma membrane / cytosol 類似検索 - 分子機能 | |||||||||
生物種 | ![]() ![]() | |||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 3.0 Å | |||||||||
![]() | Matthies D / Bae C | |||||||||
![]() | ![]() タイトル: Single-particle cryo-EM structure of a voltage-activated potassium channel in lipid nanodiscs. 著者: Doreen Matthies / Chanhyung Bae / Gilman Es Toombes / Tara Fox / Alberto Bartesaghi / Sriram Subramaniam / Kenton Jon Swartz / ![]() 要旨: Voltage-activated potassium (Kv) channels open to conduct K ions in response to membrane depolarization, and subsequently enter non-conducting states through distinct mechanisms of inactivation. X- ...Voltage-activated potassium (Kv) channels open to conduct K ions in response to membrane depolarization, and subsequently enter non-conducting states through distinct mechanisms of inactivation. X-ray structures of detergent-solubilized Kv channels appear to have captured an open state even though a non-conducting C-type inactivated state would predominate in membranes in the absence of a transmembrane voltage. However, structures for a voltage-activated ion channel in a lipid bilayer environment have not yet been reported. Here we report the structure of the Kv1.2-2.1 paddle chimera channel reconstituted into lipid nanodiscs using single-particle cryo-electron microscopy. At a resolution of ~3 Å for the cytosolic domain and ~4 Å for the transmembrane domain, the structure determined in nanodiscs is similar to the previously determined X-ray structure. Our findings show that large differences in structure between detergent and lipid bilayer environments are unlikely, and enable us to propose possible structural mechanisms for C-type inactivation. | |||||||||
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構造ビューア | EMマップ: ![]() ![]() ![]() |
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マップデータ | ![]() | 201.4 MB | ![]() | |
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ヘッダ (付随情報) | ![]() ![]() | 17.5 KB 17.5 KB | 表示 表示 | ![]() |
画像 | ![]() | 106.4 KB | ||
マスクデータ | ![]() | 216 MB | ![]() | |
Filedesc metadata | ![]() | 6.9 KB | ||
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
-検証レポート
文書・要旨 | ![]() | 586.6 KB | 表示 | ![]() |
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文書・詳細版 | ![]() | 586.2 KB | 表示 | |
XML形式データ | ![]() | 6.8 KB | 表示 | |
CIF形式データ | ![]() | 7.8 KB | 表示 | |
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「今月の分子」の関連する項目 |
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マップ
ファイル | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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注釈 | Kv1.2-2.1 paddle chimera in lipid nanodiscs, cytoplasmic domain | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
ボクセルのサイズ | X=Y=Z: 0.835 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
-マスク #1
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密度ヒストグラム |
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試料の構成要素
-全体 : Cytosolic domain of voltage-activated potassium channel Kv1.2-2.1...
全体 | 名称: Cytosolic domain of voltage-activated potassium channel Kv1.2-2.1 paddle chimera in lipid nanodiscs |
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要素 |
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-超分子 #1: Cytosolic domain of voltage-activated potassium channel Kv1.2-2.1...
超分子 | 名称: Cytosolic domain of voltage-activated potassium channel Kv1.2-2.1 paddle chimera in lipid nanodiscs タイプ: complex / ID: 1 / 親要素: 0 / 含まれる分子: #1-#2 |
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由来(天然) | 生物種: ![]() ![]() |
分子量 | 理論値: 385 KDa |
-分子 #1: Voltage-gated potassium channel subunit beta-2
分子 | 名称: Voltage-gated potassium channel subunit beta-2 / タイプ: protein_or_peptide / ID: 1 / コピー数: 4 / 光学異性体: LEVO |
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由来(天然) | 生物種: ![]() ![]() |
分子量 | 理論値: 37.339059 KDa |
組換発現 | 生物種: ![]() |
配列 | 文字列: MVQFYRNLGK SGLRVSCLGL GTWVTFGGQI TDEMAEHLMT LAYDNGINLF DTAEVYAAGK AEVVLGNIIK KKGWRRSSLV ITTKIFWGG KAETERGLSR KHIIEGLKAS LERLQLEYVD VVFANRPDPN TPMEETVRAM THVINQGMAM YWGTSRWSSM E IMEAYSVA ...文字列: MVQFYRNLGK SGLRVSCLGL GTWVTFGGQI TDEMAEHLMT LAYDNGINLF DTAEVYAAGK AEVVLGNIIK KKGWRRSSLV ITTKIFWGG KAETERGLSR KHIIEGLKAS LERLQLEYVD VVFANRPDPN TPMEETVRAM THVINQGMAM YWGTSRWSSM E IMEAYSVA RQFNLIPPIC EQAEYHMFQR EKVEVQLPEL FHKIGVGAMT WSPLACGIVS GKYDSGIPPY SRASLKGYQW LK DKILSEE GRRQQAKLKE LQAIAERLGC TLPQLAIAWC LRNEGVSSVL LGASNAEQLM ENIGAIQVLP KLSSSIVHEI DSI LGNKPY SKKDYRS UniProtKB: Voltage-gated potassium channel subunit beta-2 |
-分子 #2: Potassium voltage-gated channel subfamily A member 2,Potassium vo...
分子 | 名称: Potassium voltage-gated channel subfamily A member 2,Potassium voltage-gated channel subfamily B member 2 chimera タイプ: protein_or_peptide / ID: 2 / コピー数: 4 / 光学異性体: LEVO |
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由来(天然) | 生物種: ![]() ![]() |
分子量 | 理論値: 58.905828 KDa |
組換発現 | 生物種: ![]() |
配列 | 文字列: MAHHHHHHHH ENLYFQGSMT VATGDPVDEA AAHPGHPQDT YDPEADHECC ERVVINISGL RFETQLKTLA QFPETLLGDP KKRMRYFDP LRNEYFFDRN RPSFDAILYY YQSGGRLRRP VNVPLDIFSE EIRFYELGEE AMEMFREDEG YIKEEERPLP E NEFQRQVW ...文字列: MAHHHHHHHH ENLYFQGSMT VATGDPVDEA AAHPGHPQDT YDPEADHECC ERVVINISGL RFETQLKTLA QFPETLLGDP KKRMRYFDP LRNEYFFDRN RPSFDAILYY YQSGGRLRRP VNVPLDIFSE EIRFYELGEE AMEMFREDEG YIKEEERPLP E NEFQRQVW LLFEYPESSG PARIIAIVSV MVILISIVSF CLETLPIFRD ENEDMHGGGV TFHTYSQSTI GYQQSTSFTD PF FIVETLC IIWFSFEFLV RFFACPSKAG FFTNIMNIID IVAIIPYYVT IFLTESNKSV LQFQNVRRVV QIFRIMRILR IFK LSRHSK GLQILGQTLK ASMRELGLLI FFLFIGVILF SSAVYFAEAD ERDSQFPSIP DAFWWAVVSM TTVGYGDMVP TTIG GKIVG SLCAIAGVLT IALPVPVIVS NFNYFYHRET EGEEQAQYLQ VTSCPKIPSS PDLKKSRSAS TISKSDYMEI QEGVN NSNE DFREENLKTA NCTLANTNYV NITKMLTDV UniProtKB: Potassium voltage-gated channel subfamily A member 2, Potassium voltage-gated channel subfamily B member 2, Potassium voltage-gated channel subfamily A member 2 |
-分子 #3: NADP NICOTINAMIDE-ADENINE-DINUCLEOTIDE PHOSPHATE
分子 | 名称: NADP NICOTINAMIDE-ADENINE-DINUCLEOTIDE PHOSPHATE / タイプ: ligand / ID: 3 / コピー数: 4 / 式: NAP |
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分子量 | 理論値: 743.405 Da |
Chemical component information | ![]() ChemComp-NAP: |
-分子 #4: water
分子 | 名称: water / タイプ: ligand / ID: 4 / コピー数: 128 / 式: HOH |
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分子量 | 理論値: 18.015 Da |
Chemical component information | ![]() ChemComp-HOH: |
-実験情報
-構造解析
手法 | クライオ電子顕微鏡法 |
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![]() | 単粒子再構成法 |
試料の集合状態 | particle |
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試料調製
濃度 | 0.7 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: Quantifoil, UltrAuFoil, R1.2/1.3 / 材質: GOLD / 前処理 - タイプ: PLASMA CLEANING | ||||||||||||
凍結 | 凍結剤: ETHANE / チャンバー内湿度: 88 % / チャンバー内温度: 277.15 K / 装置: LEICA EM GP 詳細: A 3 microliter sample was applied to a plasma-cleaned grid and blotted for 10 seconds.. | ||||||||||||
詳細 | Cytosolic domain of Kv1.2-2.1 paddle chimera in lipid nanodiscs obtained after signal subtraction in RELION 2.1 |
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電子顕微鏡法
顕微鏡 | FEI TITAN KRIOS |
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撮影 | フィルム・検出器のモデル: GATAN K2 SUMMIT (4k x 4k) 検出モード: SUPER-RESOLUTION / デジタル化 - サイズ - 横: 7420 pixel / デジタル化 - サイズ - 縦: 7676 pixel / デジタル化 - 画像ごとのフレーム数: 2-20 / 撮影したグリッド数: 1 / 実像数: 3085 / 平均露光時間: 15.2 sec. / 平均電子線量: 40.0 e/Å2 |
電子線 | 加速電圧: 300 kV / 電子線源: ![]() |
電子光学系 | C2レンズ絞り径: 70.0 µm / 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): 2.5 µm / 最小 デフォーカス(公称値): 1.0 µm / 倍率(公称値): 29000 |
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 |