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基本情報
登録情報 | データベース: EMDB / ID: EMD-10375 | |||||||||
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タイトル | Complex IV in the III2-IV2 mitochondrial respiratory supercomplex from S. cerevisiae (CIV(5A)-a) | |||||||||
![]() | The reprocessed CIVa map after focused refinement with a mask including rcf2 | |||||||||
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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: 3.09 Å | |||||||||
![]() | Marechal A / Hartley A / Pinotsis N | |||||||||
資金援助 | ![]()
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![]() | ![]() タイトル: Rcf2 revealed in cryo-EM structures of hypoxic isoforms of mature mitochondrial III-IV supercomplexes. 著者: Andrew M Hartley / Brigitte Meunier / Nikos Pinotsis / Amandine Maréchal / ![]() ![]() 要旨: The organization of the mitochondrial electron transport chain proteins into supercomplexes (SCs) is now undisputed; however, their assembly process, or the role of differential expression isoforms, ...The organization of the mitochondrial electron transport chain proteins into supercomplexes (SCs) is now undisputed; however, their assembly process, or the role of differential expression isoforms, remain to be determined. In , cytochrome oxidase (CIV) forms SCs of varying stoichiometry with cytochrome (CIII). Recent studies have revealed, in normoxic growth conditions, an interface made exclusively by Cox5A, the only yeast respiratory protein that exists as one of two isoforms depending on oxygen levels. Here we present the cryo-EM structures of the III-IV and III-IV SCs containing the hypoxic isoform Cox5B solved at 3.4 and 2.8 Å, respectively. We show that the change of isoform does not affect SC formation or activity, and that SC stoichiometry is dictated by the level of CIII/CIV biosynthesis. Comparison of the CIV- and CIV-containing SC structures highlighted few differences, found mainly in the region of Cox5. Additional density was revealed in all SCs, independent of the CIV isoform, in a pocket formed by Cox1, Cox3, Cox12, and Cox13, away from the CIII-CIV interface. In the CIV-containing hypoxic SCs, this could be confidently assigned to the hypoxia-induced gene 1 (Hig1) type 2 protein Rcf2. With conserved residues in mammalian Hig1 proteins and Cox3/Cox12/Cox13 orthologs, we propose that Hig1 type 2 proteins are stoichiometric subunits of CIV, at least when within a III-IV SC. | |||||||||
履歴 |
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構造の表示
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構造ビューア | EMマップ: ![]() ![]() ![]() |
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マップデータ | ![]() | 483.6 MB | ![]() | |
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ヘッダ (付随情報) | ![]() ![]() | 23 KB 23 KB | 表示 表示 | ![]() |
FSC (解像度算出) | ![]() | 17.8 KB | 表示 | ![]() |
画像 | ![]() | 110.6 KB | ||
アーカイブディレクトリ | ![]() ![]() | HTTPS FTP |
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リンク
EMDBのページ | ![]() ![]() |
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マップ
ファイル | ![]() | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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注釈 | The reprocessed CIVa map after focused refinement with a mask including rcf2 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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: 1.048 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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試料の構成要素
+全体 : Complex IV in the III2-IV2 mitochondrial respiratory supercomplex...
+超分子 #1: Complex IV in the III2-IV2 mitochondrial respiratory supercomplex...
+超分子 #2: Complex IV in the III2-IV2 mitochondrial respiratory supercomplex...
+超分子 #3: Complex IV in the III2-IV2 mitochondrial respiratory supercomplex...
+分子 #1: CYTOCHROME C OXIDASE SUBUNIT 1; SYNONYM: CYTOCHROME C OXIDASE POL...
+分子 #2: CYTOCHROME C OXIDASE SUBUNIT 2; SYNONYM: CYTOCHROME C OXIDASE POL...
+分子 #3: CYTOCHROME C OXIDASE SUBUNIT 3; SYNONYM: CYTOCHROME C OXIDASE POL...
+分子 #4: CYTOCHROME C OXIDASE SUBUNIT 4, MITOCHONDRIAL; SYNONYM: CYTOCHROM...
+分子 #5: CYTOCHROME C OXIDASE POLYPEPTIDE 5A, MITOCHONDRIAL; SYNONYM: CYTO...
+分子 #6: CYTOCHROME C OXIDASE SUBUNIT 6, MITOCHONDRIAL; SYNONYM: CYTOCHROM...
+分子 #7: CYTOCHROME C OXIDASE SUBUNIT 7; SYNONYM: CYTOCHROME C OXIDASE POL...
+分子 #8: CYTOCHROME C OXIDASE POLYPEPTIDE VIII, MITOCHONDRIAL; SYNONYM: COX8
+分子 #9: CYTOCHROME C OXIDASE SUBUNIT 7A; SYNONYM: CYTOCHROME C OXIDASE PO...
+分子 #10: CYTOCHROME C OXIDASE SUBUNIT 6B; SYNONYM: CYTOCHROME C OXIDASE PO...
+分子 #11: CYTOCHROME C OXIDASE SUBUNIT 6A, MITOCHONDRIAL; CYTOCHROME C OXID...
+分子 #12: COX26; SYNONYM: Uncharacterized protein YDR119W-A
+分子 #13: RCF2; SYNONYM: Respiratory supercomplex factor 2
-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
緩衝液 | pH: 7.2 |
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凍結 | 凍結剤: ETHANE / 装置: FEI VITROBOT MARK IV 詳細: 3 uL of sample applied to negatively glow discharged grid, blot force -10; blotting time 8.5 sec.. |
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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) 実像数: 2740 / 平均電子線量: 52.64 e/Å2 |
電子線 | 加速電圧: 300 kV / 電子線源: ![]() |
電子光学系 | 照射モード: FLOOD BEAM / 撮影モード: BRIGHT FIELD |
実験機器 | ![]() モデル: Titan Krios / 画像提供: FEI Company |