How many molecules of mitochondrial complex I are in a cell?

How many molecules of mitochondrial complex I are in a cell?
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DOI:
10.1016/j.ab.2022.114646
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发表时间:
2022-06-01
影响因子:
2.9
通讯作者:
--
中科院分区:
生物学4区
文献类型:
--
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线粒体复合物 I 是唯一负责基质 NADH 氧化和 NAD+ 分解代谢再生的酶。核和线粒体DNA突变、组装损伤和酶损伤与遗传性疾病、缺血再灌注损伤、神经退行性变和肿瘤发生有关。在这里,我们介绍一种测量复合物 I 绝对含量的新方法。该方法基于通过 Clear Native 电泳分离复合物后对聚丙烯酰胺凝胶进行黄素荧光扫描。以小鼠原代星形胶质细胞为例,我们计算出2.2×105个复合物I分子/细胞的平均值。我们的方法可用于准确定量复合物 I 含量。
Mitochondrial complex I is the only enzyme responsible for oxidation of matrix NADH and regeneration of NAD+ for catabolism. Nuclear and mtDNA mutations, assembly impairments, and enzyme damage are implicated in inherited diseases, ischemia-reperfusion injury, neurodegeneration, and tumorogenesis. Here we introduce a novel method to measure the absolute content of complex I. The method is based on flavin fluorescence scanning of a polyacrylamide gel after separation of complexes by Clear Native electrophoresis. Using mouse primary astrocytes as an example, we calculated an average value of 2.2×105 complex I molecules/cell. Our method can be used for accurate quantification of complex I content.
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