+データを開く
-基本情報
登録情報 | データベース: EMDB / ID: EMD-16962 | |||||||||
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タイトル | Mitochondrial complex I from Mus musculus in the active state bound with piericidin A | |||||||||
マップデータ | Full map of mouse complex I inhibited by Piericidin A, post-processed with RELION | |||||||||
試料 |
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キーワード | Mitochondrial complex I / Respiratory complex I / NADH:ubiquinone oxidoreductase / OXIDOREDUCTASE | |||||||||
機能・相同性 | 機能・相同性情報 Mitochondrial protein import / response to injury involved in regulation of muscle adaptation / reproductive system development / Protein lipoylation / Mitochondrial Fatty Acid Beta-Oxidation / Complex I biogenesis / RHOG GTPase cycle / Respiratory electron transport / protein insertion into mitochondrial inner membrane / blastocyst hatching ...Mitochondrial protein import / response to injury involved in regulation of muscle adaptation / reproductive system development / Protein lipoylation / Mitochondrial Fatty Acid Beta-Oxidation / Complex I biogenesis / RHOG GTPase cycle / Respiratory electron transport / protein insertion into mitochondrial inner membrane / blastocyst hatching / circulatory system development / respiratory system process / psychomotor behavior / ubiquinone-6 biosynthetic process / Mitochondrial protein degradation / response to light intensity / cellular response to oxygen levels / iron-sulfur cluster assembly complex / : / mitochondrial large ribosomal subunit binding / gliogenesis / negative regulation of non-canonical NF-kappaB signal transduction / neural precursor cell proliferation / cardiac muscle tissue development / [2Fe-2S] cluster assembly / oxygen sensor activity / adult walking behavior / positive regulation of mitochondrial membrane potential / cellular response to glucocorticoid stimulus / response to hydroperoxide / : / ubiquinone binding / iron-sulfur cluster assembly / positive regulation of ATP biosynthetic process / adult behavior / dopamine metabolic process / NADH:ubiquinone reductase (H+-translocating) / positive regulation of execution phase of apoptosis / proton motive force-driven mitochondrial ATP synthesis / apoptotic mitochondrial changes / NADH dehydrogenase activity / mitochondrial ATP synthesis coupled electron transport / : / mitochondrial respiratory chain complex I assembly / mitochondrial electron transport, NADH to ubiquinone / electron transport coupled proton transport / respiratory chain complex I / acyl binding / acyl carrier activity / NADH dehydrogenase (ubiquinone) activity / cellular response to interferon-beta / neuron development / negative regulation of intrinsic apoptotic signaling pathway / ATP synthesis coupled electron transport / quinone binding / negative regulation of reactive oxygen species biosynthetic process / cellular response to retinoic acid / extrinsic apoptotic signaling pathway / muscle contraction / tricarboxylic acid cycle / ionotropic glutamate receptor binding / Neutrophil degranulation / aerobic respiration / response to hormone / visual perception / respiratory electron transport chain / cerebellum development / reactive oxygen species metabolic process / neurogenesis / regulation of mitochondrial membrane potential / kidney development / mitochondrion organization / fatty acid metabolic process / synaptic membrane / response to cocaine / response to nicotine / mitochondrial membrane / sensory perception of sound / electron transport chain / regulation of protein phosphorylation / brain development / negative regulation of cell growth / multicellular organism growth / response to hydrogen peroxide / response to organic cyclic compound / mitochondrial intermembrane space / cognition / 2 iron, 2 sulfur cluster binding / circadian rhythm / positive regulation of protein catabolic process / NAD binding / FMN binding / myelin sheath / nervous system development / 4 iron, 4 sulfur cluster binding / protease binding / neuron apoptotic process / response to ethanol / in utero embryonic development / response to oxidative stress 類似検索 - 分子機能 | |||||||||
生物種 | Mus musculus (ハツカネズミ) / house mouse (ハツカネズミ) | |||||||||
手法 | 単粒子再構成法 / クライオ電子顕微鏡法 / 解像度: 2.84 Å | |||||||||
データ登録者 | Grba DN / Chung I / Bridges HR / Agip ANA / Hirst J | |||||||||
資金援助 | 英国, 2件
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引用 | ジャーナル: Sci Adv / 年: 2023 タイトル: Investigation of hydrated channels and proton pathways in a high-resolution cryo-EM structure of mammalian complex I. 著者: Daniel N Grba / Injae Chung / Hannah R Bridges / Ahmed-Noor A Agip / Judy Hirst / 要旨: Respiratory complex I, a key enzyme in mammalian metabolism, captures the energy released by reduction of ubiquinone by NADH to drive protons across the inner mitochondrial membrane, generating the ...Respiratory complex I, a key enzyme in mammalian metabolism, captures the energy released by reduction of ubiquinone by NADH to drive protons across the inner mitochondrial membrane, generating the proton-motive force for ATP synthesis. Despite remarkable advances in structural knowledge of this complicated membrane-bound enzyme, its mechanism of catalysis remains controversial. In particular, how ubiquinone reduction is coupled to proton pumping and the pathways and mechanisms of proton translocation are contested. We present a 2.4-Å resolution cryo-EM structure of complex I from mouse heart mitochondria in the closed, active (ready-to-go) resting state, with 2945 water molecules modeled. By analyzing the networks of charged and polar residues and water molecules present, we evaluate candidate pathways for proton transfer through the enzyme, for the chemical protons for ubiquinone reduction, and for the protons transported across the membrane. Last, we compare our data to the predictions of extant mechanistic models, and identify key questions to answer in future work to test them. | |||||||||
履歴 |
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-構造の表示
添付画像 |
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-ダウンロードとリンク
-EMDBアーカイブ
マップデータ | emd_16962.map.gz | 457.7 MB | EMDBマップデータ形式 | |
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ヘッダ (付随情報) | emd-16962-v30.xml emd-16962.xml | 72.5 KB 72.5 KB | 表示 表示 | EMDBヘッダ |
FSC (解像度算出) | emd_16962_fsc.xml | 17.7 KB | 表示 | FSCデータファイル |
画像 | emd_16962.png | 104.7 KB | ||
マスクデータ | emd_16962_msk_1.map | 488.4 MB | マスクマップ | |
その他 | emd_16962_half_map_1.map.gz emd_16962_half_map_2.map.gz | 394.3 MB 394.8 MB | ||
アーカイブディレクトリ | http://ftp.pdbj.org/pub/emdb/structures/EMD-16962 ftp://ftp.pdbj.org/pub/emdb/structures/EMD-16962 | HTTPS FTP |
-検証レポート
文書・要旨 | emd_16962_validation.pdf.gz | 1.1 MB | 表示 | EMDB検証レポート |
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文書・詳細版 | emd_16962_full_validation.pdf.gz | 1.1 MB | 表示 | |
XML形式データ | emd_16962_validation.xml.gz | 25.6 KB | 表示 | |
CIF形式データ | emd_16962_validation.cif.gz | 34.2 KB | 表示 | |
アーカイブディレクトリ | https://ftp.pdbj.org/pub/emdb/validation_reports/EMD-16962 ftp://ftp.pdbj.org/pub/emdb/validation_reports/EMD-16962 | HTTPS FTP |
-関連構造データ
関連構造データ | 8oltMC 8om1C M: このマップから作成された原子モデル C: 同じ文献を引用 (文献) |
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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
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_16962.map.gz / 形式: CCP4 / 大きさ: 488.4 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES) | ||||||||||||||||||||||||||||||||||||
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注釈 | Full map of mouse complex I inhibited by Piericidin A, post-processed with RELION | ||||||||||||||||||||||||||||||||||||
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: 1.0625 Å | ||||||||||||||||||||||||||||||||||||
密度 |
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対称性 | 空間群: 1 | ||||||||||||||||||||||||||||||||||||
詳細 | EMDB XML:
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-添付データ
-マスク #1
ファイル | emd_16962_msk_1.map | ||||||||||||
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投影像・断面図 |
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密度ヒストグラム |
-ハーフマップ: Half map 2
ファイル | emd_16962_half_map_1.map | ||||||||||||
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注釈 | Half map 2 | ||||||||||||
投影像・断面図 |
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密度ヒストグラム |
-ハーフマップ: Half map 1
ファイル | emd_16962_half_map_2.map | ||||||||||||
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注釈 | Half map 1 | ||||||||||||
投影像・断面図 |
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密度ヒストグラム |
-試料の構成要素
+全体 : Mitochondrial respiratory complex I
+超分子 #1: Mitochondrial respiratory complex I
+分子 #1: NADH-ubiquinone oxidoreductase chain 3
+分子 #2: NADH dehydrogenase [ubiquinone] iron-sulfur protein 7, mitochondrial
+分子 #3: NADH dehydrogenase [ubiquinone] iron-sulfur protein 3, mitochondrial
+分子 #4: NADH dehydrogenase [ubiquinone] iron-sulfur protein 2, mitochondrial
+分子 #5: NADH dehydrogenase [ubiquinone] flavoprotein 2, mitochondrial
+分子 #6: NADH dehydrogenase [ubiquinone] flavoprotein 1, mitochondrial
+分子 #7: NADH-ubiquinone oxidoreductase 75 kDa subunit, mitochondrial
+分子 #8: NADH-ubiquinone oxidoreductase chain 1
+分子 #9: NADH dehydrogenase [ubiquinone] iron-sulfur protein 8, mitochondrial
+分子 #10: NADH-ubiquinone oxidoreductase chain 6
+分子 #11: NADH-ubiquinone oxidoreductase chain 4L
+分子 #12: NADH-ubiquinone oxidoreductase chain 5
+分子 #13: NADH-ubiquinone oxidoreductase chain 4
+分子 #14: NADH-ubiquinone oxidoreductase chain 2
+分子 #15: NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 10, mi...
+分子 #16: NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 9, mit...
+分子 #17: NADH dehydrogenase [ubiquinone] iron-sulfur protein 4, mitochondrial
+分子 #18: NADH dehydrogenase [ubiquinone] iron-sulfur protein 6, mitochondrial
+分子 #19: NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 2
+分子 #20: Acyl carrier protein, mitochondrial
+分子 #21: NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 5
+分子 #22: NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 6
+分子 #23: NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 8
+分子 #24: NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 11
+分子 #25: NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 13
+分子 #26: NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 1
+分子 #27: NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 3
+分子 #28: NADH dehydrogenase [ubiquinone] 1 subunit C1, mitochondrial
+分子 #29: NADH dehydrogenase [ubiquinone] 1 subunit C2
+分子 #30: NADH dehydrogenase [ubiquinone] iron-sulfur protein 5
+分子 #31: NADH dehydrogenase [ubiquinone] 1 beta subcomplex subunit 1
+分子 #32: NADH dehydrogenase [ubiquinone] 1 beta subcomplex subunit 11, mit...
+分子 #33: NADH dehydrogenase [ubiquinone] 1 beta subcomplex subunit 5, mito...
+分子 #34: NADH dehydrogenase [ubiquinone] 1 beta subcomplex subunit 6
+分子 #35: NADH dehydrogenase [ubiquinone] 1 beta subcomplex subunit 2, mito...
+分子 #36: NADH dehydrogenase [ubiquinone] 1 beta subcomplex subunit 3
+分子 #37: NADH dehydrogenase [ubiquinone] 1 beta subcomplex subunit 8, mito...
+分子 #38: NADH dehydrogenase [ubiquinone] 1 beta subcomplex subunit 4
+分子 #39: NADH dehydrogenase [ubiquinone] 1 beta subcomplex subunit 9
+分子 #40: NADH dehydrogenase [ubiquinone] 1 beta subcomplex subunit 7, METH...
+分子 #41: NADH dehydrogenase [ubiquinone] 1 beta subcomplex subunit 10
+分子 #42: NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 12
+分子 #43: NADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 7
+分子 #44: NADH dehydrogenase [ubiquinone] flavoprotein 3, mitochondrial
+分子 #45: DODECYL-BETA-D-MALTOSIDE
+分子 #46: 1,2-DIACYL-SN-GLYCERO-3-PHOSPHOCHOLINE
+分子 #47: IRON/SULFUR CLUSTER
+分子 #48: Piericidin A
+分子 #49: FE2/S2 (INORGANIC) CLUSTER
+分子 #50: FLAVIN MONONUCLEOTIDE
+分子 #51: SODIUM ION
+分子 #52: CARDIOLIPIN
+分子 #53: 1,2-Distearoyl-sn-glycerophosphoethanolamine
+分子 #54: MAGNESIUM ION
+分子 #55: GUANOSINE-5'-TRIPHOSPHATE
+分子 #56: NADPH DIHYDRO-NICOTINAMIDE-ADENINE-DINUCLEOTIDE PHOSPHATE
+分子 #57: ZINC ION
+分子 #58: ~{S}-[2-[3-[[(2~{R})-3,3-dimethyl-2-oxidanyl-4-phosphonooxy-butan...
+分子 #59: MYRISTIC ACID
+分子 #60: water
-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 | 4.1 mg/mL | ||||||||||||
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緩衝液 | pH: 7.14 構成要素:
詳細: PH corrected at room temperature | ||||||||||||
グリッド | モデル: UltrAuFoil / 材質: GOLD / メッシュ: 300 / 支持フィルム - 材質: GOLD / 支持フィルム - トポロジー: HOLEY / 前処理 - タイプ: GLOW DISCHARGE / 前処理 - 時間: 90 sec. / 前処理 - 雰囲気: AIR 詳細: The grid was treated for 48 hours in an anaerobic glovebox in ethanol containing 5mM 11-mercaptoundecylhexaethyleneglycol, washed three times in ethanol and air dried prior to use. Note the hole sizes are R0.6/1. | ||||||||||||
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: FEI VITROBOT MARK IV / 詳細: blot for 10 to 12 seconds before plunging. |
-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 Quantum LS / エネルギーフィルター - スリット幅: 20 eV |
撮影 | #0 - Image recording ID: 1 #0 - フィルム・検出器のモデル: GATAN K2 SUMMIT (4k x 4k) #0 - 検出モード: COUNTING / #0 - デジタル化 - サイズ - 横: 3838 pixel / #0 - デジタル化 - サイズ - 縦: 3710 pixel / #0 - デジタル化 - 画像ごとのフレーム数: 1-21 / #0 - 撮影したグリッド数: 1 / #0 - 実像数: 1200 / #0 - 平均露光時間: 10.0 sec. / #0 - 平均電子線量: 50.0 e/Å2 #0 - 詳細: Images were collected over 25 frames per exposure #1 - Image recording ID: 2 #1 - フィルム・検出器のモデル: FEI FALCON III (4k x 4k) #1 - 検出モード: COUNTING / #1 - 撮影したグリッド数: 1 / #1 - 実像数: 1454 / #1 - 平均露光時間: 71.5 sec. / #1 - 平均電子線量: 46.0 e/Å2 #1 - 詳細: Images were collected over 40 frames per exposure |
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): 3.4 µm / 最小 デフォーカス(公称値): 2.2 µm / 倍率(公称値): 47600 |
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 |