ジャーナル: EMBO J / 年: 2011 タイトル: Arrangement of electron transport chain components in bovine mitochondrial supercomplex I1III2IV1. 著者: Thorsten Althoff / Deryck J Mills / Jean-Luc Popot / Werner Kühlbrandt / 要旨: The respiratory chain in the inner mitochondrial membrane contains three large multi-enzyme complexes that together establish the proton gradient for ATP synthesis, and assemble into a supercomplex. ...The respiratory chain in the inner mitochondrial membrane contains three large multi-enzyme complexes that together establish the proton gradient for ATP synthesis, and assemble into a supercomplex. A 19-Å 3D map of the 1.7-MDa amphipol-solubilized supercomplex I(1)III(2)IV(1) from bovine heart obtained by single-particle electron cryo-microscopy reveals an amphipol belt replacing the membrane lipid bilayer. A precise fit of the X-ray structures of complex I, the complex III dimer, and monomeric complex IV indicates distances of 13 nm between the ubiquinol-binding sites of complexes I and III, and of 10-11 nm between the cytochrome c binding sites of complexes III and IV. The arrangement of respiratory chain complexes suggests two possible pathways for efficient electron transfer through the supercomplex, of which the shorter branch through the complex III monomer proximal to complex I may be preferred.
#1: 年: 2011 タイトル: Strukturelle Untersuchungen am Superkomplex I1III2IV1 der Atmungskette mittels Kryoelektronenmikroskopie 著者: Althoff T
#241 - 2020年1月 20年の分子を振り返って (Twenty Years of Molecules) 類似性 (8)
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マップ
ファイル
ダウンロード / ファイル: emd_1876.map.gz / 形式: CCP4 / 大きさ: 5.2 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
注釈
3D cryo-EM map of supercomplex B (I1III2IV1) from bovine heart mitochondria
ボクセルのサイズ
X=Y=Z: 4.76 Å
密度
表面レベル
登録者による: 1.3 / ムービー #1: 1.3
最小 - 最大
-8.248889999999999 - 18.839099999999998
平均 (標準偏差)
-0.00000000106836 (±1.0)
対称性
空間群: 1
詳細
EMDB XML:
マップ形状
Axis order
Y
X
Z
Origin
-56
-56
-55
サイズ
112
112
112
Spacing
112
112
112
セル
A=B=C: 533.12 Å α=β=γ: 90 °
CCP4マップ ヘッダ情報:
mode
Image stored as Reals
Å/pix. X/Y/Z
4.76
4.76
4.76
M x/y/z
112
112
112
origin x/y/z
0.000
0.000
0.000
length x/y/z
533.120
533.120
533.120
α/β/γ
90.000
90.000
90.000
start NX/NY/NZ
-56
-56
-55
NX/NY/NZ
112
112
112
MAP C/R/S
2
1
3
start NC/NR/NS
-56
-56
-55
NC/NR/NS
112
112
112
D min/max/mean
-8.249
18.839
-0.000
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添付データ
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試料の構成要素
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全体 : Respiratory supercomplex B (I1III2IV1) composed of complex I, dim...
全体
名称: Respiratory supercomplex B (I1III2IV1) composed of complex I, dimeric complex III and complex IV from bovine heart mitochondria
要素
試料: Respiratory supercomplex B (I1III2IV1) composed of complex I, dimeric complex III and complex IV from bovine heart mitochondria
タンパク質・ペプチド: NADH-dehydrogenase
タンパク質・ペプチド: Cytochrome c reductase
タンパク質・ペプチド: Cytochrome c oxidase
タンパク質・ペプチド: Cytochrome c
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超分子 #1000: Respiratory supercomplex B (I1III2IV1) composed of complex I, dim...
超分子
名称: Respiratory supercomplex B (I1III2IV1) composed of complex I, dimeric complex III and complex IV from bovine heart mitochondria タイプ: sample / ID: 1000 詳細: The supercomplexes were kept soluble by amphipol A8-35 集合状態: A monomer of complex I assembles with a dimer of complex III and a monomer of complex IV Number unique components: 4
pH: 7.7 / 詳細: 10 mM KCl, 15 mM HEPES, residual trehalose
グリッド
詳細: Continuous carbon 400 mesh copper grid
凍結
凍結剤: ETHANE / チャンバー内湿度: 70 % / チャンバー内温度: 95 K / 装置: HOMEMADE PLUNGER 詳細: Vitrification instrument: Custom-made hand plunger. Sample was adsorbed to carbon film for 30 seconds before blotting 手法: One-sided blotting for 26 - 30 seconds before plunging
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電子顕微鏡法
顕微鏡
FEI POLARA 300
温度
最低: 78 K / 最高: 95 K / 平均: 78 K
アライメント法
Legacy - 非点収差: Objective lens astigmatism was corrected at 200,000 x magnification Legacy - Electron beam tilt params: Beam tilt was corrected by adjusting pivot points
詳細
Low dose conditions. For random conical tilt each area was imaged twice with the first image taken at a tilt angle of -45 or -50 degree
撮影
カテゴリ: FILM / フィルム・検出器のモデル: KODAK SO-163 FILM / デジタル化 - スキャナー: ZEISS SCAI / デジタル化 - サンプリング間隔: 7 µm / 実像数: 154 / 平均電子線量: 25 e/Å2 詳細: Scanned images were binned to 2.38 Angstroms per pixel in IMAGIC Od range: 0.7 / ビット/ピクセル: 8
Tilt pairs of particles were selected interactively in JWEB
CTF補正
詳細: Phase-flippping on each particle
最終 再構成
想定した対称性 - 点群: C1 (非対称) / アルゴリズム: OTHER / 解像度のタイプ: BY AUTHOR / 解像度: 19.0 Å / 解像度の算出法: FSC 0.143 CUT-OFF / ソフトウェア - 名称: SPIDER, IMAGIC 詳細: Final map was calculated from combined tilted and untilted datasets with 10684 projections each and omitting 30 per cent of images with lowest cross correlation coefficient using SIRT 使用した粒子像数: 10684
Protocol: Rigid Body. Only one monomer from the asymmetric unit was used. Docking was done manually according to detailed map features. Gave better fit than
精密化
空間: REAL / プロトコル: RIGID BODY FIT
得られたモデル
PDB-2ybb: Fitted model for bovine mitochondrial supercomplex I1III2IV1 by single particle cryo-EM (EMD-1876)