Method for detecting CoQ10 incorporation in the mitochondrial respiratory chain supercomplex.

Method for detecting CoQ10 incorporation in the mitochondrial respiratory chain supercomplex.
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DOI:
10.3164/jcbn.22-137
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发表时间:
2023-05
影响因子:
2.4
通讯作者:
--
中科院分区:
医学4区
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--
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辅酶Q10是线粒体电子传递链的重要组成部分。线粒体电子传递系统蛋白的超复合物存在。该复合物还含有辅酶Q10。辅酶Q10在组织中的浓度随年龄和病理而降低。辅酶Q10作为补充剂。目前还不清楚辅酶Q10是否被转运到超复合物中。本研究建立了一种测定线粒体呼吸链超复合物中辅酶Q10的方法。蓝色天然电泳用于分离线粒体膜。将电泳凝胶切成3 mm切片。 用正己烷提取该切片中的辅酶Q10,用HPLC-ECD分析辅酶Q10。辅酶Q10在凝胶中与超复合物的位置相同。在这个位置的辅酶Q10被认为是超复合物中的辅酶Q10。我们发现,4-硝基苯甲酸,辅酶Q10的生物合成抑制剂,减少辅酶Q10的超复合物内外的量。我们还观察到,向细胞中加入辅酶Q10增加了超复合物中辅酶Q10的量。该方法有望用于分析各种样品中超复合物中辅酶Q10的含量。
Coenzyme Q10 is an important component of the mitochondrial electron transfer chain. A supercomplex of mitochondrial electron transfer system proteins exists. This complex also contains coenzyme Q10. The concentrations of coenzyme Q10 in tissues decrease with age and pathology. Coenzyme Q10 is given as a supplement. It is unknown whether coenzyme Q10 is transported to the supercomplex. We develop a method for measuring coenzyme Q10 in the mitochondrial respiratory chain supercomplex in this study. Blue native electrophoresis was used to separate mitochondrial membranes. Electrophoresis gels were cut into 3 mm slices. Hexane was used to extract coenzyme Q10 from this slice, and HPLC-ECD was used to analyze coenzyme Q10. Coenzyme Q10 was found in the gel at the same site as the supercomplex. Coenzyme Q10 at this location was thought to be coenzyme Q10 in the supercomplex. We discovered that 4-nitrobenzoate, a coenzyme Q10 biosynthesis inhibitor, reduced the amount of coenzyme Q10 both within and outside the supercomplex. We also observed that the addition of coenzyme Q10 to cells increased the amount of coenzyme Q10 in the supercomplex. It is expected to analysis coenzyme Q10 level in supercomplex in various samples by using this novel method.
DOI: 10.1242/jcs.258399
发表时间: 2021-07-01
影响因子: 4
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发表时间: 1989-07-01
期刊: LIPIDS
影响因子: 1.9
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通讯作者: DALLNER, G
DOI: 10.1073/pnas.82.13.4513
发表时间: 1985-01-01
影响因子: 11.1
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