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[通过共价连接的pH探针检测细胞内线粒体膜上H⁺离子的局部梯度及其功能作用]

[Detection and functional role of local gradients of H+-ions on the intracellular mitochondrial membrane with covalently linked pH-probe].

作者信息

Kozlova M V, Gramadskiĭ K B, Solodovnikova I M, Krasinskaia I P, Vinogradov A V, Iaguzhinskiĭ L S

机构信息

Belozerskii Institute of Physicochemical Biology, Lomonosov Moscow State University, Vorob'evy Gory, Moscow, 119889 Russia.

出版信息

Biofizika. 2003 May-Jun;48(3):443-52.

Abstract

The study is devoted to the registration of local H+ gradients on the inner membrane of mitochondria under conditions of H+ pump functioning were recorded. By using a covalently linked pH probe (fluorescein isothiocyanate), a local increase in the activity of hydrogen ions on the outer face of the inner mitochondrial membrane in the presence of the respiration substrate at increased permeability of the membrane for K+ was registered. It was also found that the buffer capacity of medium affects the respiration rate of completely uncoupled mitochondria; a change in respiration rate strictly correlates with changes in local H+ gradients on the mitochondrial membrane. It was concluded that local gradients of H+ activity can control the rate of functioning of H+ pumps. It was shown that, under certain conditions, the system of H+ pumps incorporated into succinate oxidase of mitochondria functions as a nonliner system.

摘要

该研究致力于记录在H⁺泵功能条件下线粒体内膜上局部H⁺梯度。通过使用共价连接的pH探针(异硫氰酸荧光素),在呼吸底物存在且膜对K⁺通透性增加的情况下,记录到线粒体内膜外表面氢离子活性局部增加。还发现培养基的缓冲能力会影响完全解偶联线粒体的呼吸速率;呼吸速率的变化与线粒体膜上局部H⁺梯度的变化严格相关。得出的结论是,H⁺活性的局部梯度可以控制H⁺泵的功能速率。结果表明,在某些条件下,整合到线粒体琥珀酸氧化酶中的H⁺泵系统作为一个非线性系统起作用。

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