Cysteine scanning reveals minor local rearrangements of the horizontal helix of respiratory complex I

Cysteine scanning reveals minor local rearrangements of the horizontal helix of respiratory complex I
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半胱氨酸扫描显示呼吸复合物 I 水平螺旋的微小局部重排

DOI:
10.1111/mmi.13112
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发表时间:
2015
影响因子:
3.6
通讯作者:
Friedrich
Friedrich
中科院分区:
生物学2区
文献类型:
--
作者:
Steimle;Schnick;Burger;Krämer;Dawitz;Brander;Matlosz;Schäfer;Maurer;Glessner;Friedrich

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NADH:泛醌氧化还原酶,呼吸复合物I,将电子从NADH转移到泛醌与质子跨膜易位偶联。该配合物由催化氧化还原反应的外周臂和催化质子移位的膜臂组成。膜臂几乎完全由110 μ m独特的水平螺旋排列,该螺旋被讨论为将由氧化还原反应引起的构象变化以活塞样运动传递到膜臂,从而驱动质子移位。在这里,我们通过半胱氨酸扫描大肠杆菌复合物I的螺旋来分析这种拟议的运动。通过在氧化和还原状态下分别用荧光标记物和自旋探针标记制备物,测定工程化半胱氨酸残基的可及性和单个位置的灵活性。两种氧化还原状态之间的自旋探针的荧光标记和旋转灵活性的差异表明在螺旋的不同位置只有轻微的构象变化,但不是一个大的运动。
The NADH:ubiquinone oxidoreductase, respiratory complex I, couples electron transfer from NADH to ubiquinone with the translocation of protons across the membrane. The complex consists of a peripheral arm catalyzing the redox reaction and a membrane arm catalyzing proton translocation. The membrane arm is almost completely aligned by a 110 Å unique horizontal helix that is discussed to transmit conformational changes induced by the redox reaction in a piston‐like movement to the membrane arm driving proton translocation. Here, we analyzed such a proposed movement by cysteine‐scanning of the helix of theEscherichia colicomplex I. The accessibility of engineered cysteine residues and the flexibility of individual positions were determined by labeling the preparations with a fluorescent marker and a spin‐probe, respectively, in the oxidized and reduced states. The differences in fluorescence labeling and the rotational flexibility of the spin probe between both redox states indicate only slight conformational changes at distinct positions of the helix but not a large movement.
呼吸复合体 I 的结构视角
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