低需氧能力海洋无脊椎动物透化纤维中线粒体呼吸的原位定量
In situ quantification of mitochondrial respiration in permeabilized fibers of a marine invertebrate with low aerobic capacity
Keywords:Alternative oxidase;Marine invertebrates;Metabolism;Mitochondrial respiration;Permeabilized fibers
关键词:交替氧化酶;海洋无脊椎动物;新陈代谢;线粒体呼吸;透化纤维
非哺乳动物:环节动物
作者:Pichaud N, Rioux P, Blier PU
出版期刊:《Comparative Biochemistry and Physiology Part A: Molecular & Integrative Physiology》 2012/1/8
Abstract:
The aim of this study was to design a protocol to allow the assessment of normal and alternative pathways for electron transport in mitochondria using an in situ approach (on permeabilized fibers) in high-resolution respirometry. We measured the oxygen consumption of permeabilized fibers from Nereis (Neanthes) virens with different substrates and the presence of ADP. To estimate the alternative oxidase (AOX) activity, antimycin A was introduced in order to inhibit complex III. Moreover, the apparent complex IV (COX) excess capacity was evaluated using different substrates to assess the implication of this complex in the partitioning of electrons during its progressive inhibition. Our in situ method enabled to quantify the activity of the normal COX pathway as well as the AOX pathway when different substrates were oxidized by either complex I, complex II or both. Using this approach, we confirmed that according to the substrates used, each pathway has a different role and consequently is otherwise involved in the partitioning of electrons through the electron transport system, and suggested that the AOX activity is triggered not only by the redox state of the cell but also by the type of substrates provided to mitochondria.
文章摘要:
本研究的目的是设计一个协议,以允许在高分辨率呼吸测量中使用原位方法(在透化纤维上)评估线粒体中电子传输的正常和替代途径。我们测量了来自 Nereis (Neanthes) virens 的透化纤维在不同底物和 ADP 存在下的耗氧量。为了估计替代氧化酶 (AOX) 活性,引入抗霉素 A 以抑制复合物 III。此外,使用不同的底物评估表观复合物 IV (COX) 过剩容量,以评估这种复合物在其渐进抑制过程中对电子分配的影响。当不同的底物被复合物 I、复合物 II 或两者氧化时,我们的原位方法能够量化正常 COX 途径和 AOX 途径的活性。使用这种方法,我们证实,根据所使用的底物,每条途径都有不同的作用,因此参与电子通过电子传输系统的分配,并表明 AOX 活性不仅由氧化还原状态触发。细胞还由提供给线粒体的底物类型决定。
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文章题目、关键词与摘要译文仅用于参考。