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

IF 2 4区 医学 Q3 NUTRITION & DIETETICS Journal of Clinical Biochemistry and Nutrition Pub Date : 2023-05-01 DOI:10.3164/jcbn.22-137
Kyousuke Sugawara, Seiji Sato, Yuto Tanaka, Akari Nakamura, Akio Fujisawa, Yorihiro Yamamoto, Misato Kashiba
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Abstract

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.

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线粒体呼吸链超复合体中辅酶q10掺入的检测方法。
辅酶Q10是线粒体电子传递链的重要组成部分。存在线粒体电子传递系统蛋白的超复合体。这个复合体也含有辅酶Q10。组织中辅酶Q10的浓度随年龄和病理变化而降低。辅酶Q10作为补充剂。目前尚不清楚辅酶Q10是否被转运到超复合体中。在这项研究中,我们开发了一种测量线粒体呼吸链超复合体中辅酶Q10的方法。采用蓝色原生电泳分离线粒体膜。将电泳凝胶切成3毫米的薄片。用己烷提取该切片中的辅酶Q10,用HPLC-ECD分析辅酶Q10。在凝胶中发现辅酶Q10位于超络合物的同一位置。这个位置的辅酶Q10被认为是超复合体中的辅酶Q10。我们发现辅酶Q10生物合成抑制剂4-硝基苯甲酸酯可以减少超络合物内外辅酶Q10的量。我们还观察到,在细胞中添加辅酶Q10增加了超复合体中辅酶Q10的数量。该方法有望用于各种样品超络合物中辅酶Q10水平的分析。
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来源期刊
CiteScore
4.30
自引率
8.30%
发文量
57
审稿时长
6-12 weeks
期刊介绍: Journal of Clinical Biochemistry and Nutrition (JCBN) is an international, interdisciplinary publication encompassing chemical, biochemical, physiological, pathological, toxicological and medical approaches to research on lipid peroxidation, free radicals, oxidative stress and nutrition. The Journal welcomes original contributions dealing with all aspects of clinical biochemistry and clinical nutrition including both in vitro and in vivo studies.
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