An improved spectrophotometric method for a more specific and accurate assay of mitochondrial complex III activity
An improved spectrophotometric method for a more specific and accurate assay of mitochondrial complex III activity
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DOI：
10.1016/j.cca.2008.04.025
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发表时间：
2008-09-01
影响因子：
5
通讯作者：
Liu, Jiankang
中科院分区：
文献类型：
--
作者：
Luo, Cheng;Long, Jiangang;Liu, Jiankang
背景：组织样本中复合体III活性的测定为线粒体疾病和与线粒体功能障碍相关的疾病的诊断提供了关键证据。方法：研究了一种比现有方法具有更高特异性的改良分光光度法的反应条件。结果：确定了最佳反应条件：牛血清白蛋白为0.01%，吐温-20为0.05%。与以往的方法相比，本方法具有更强的抗霉素A敏感复合体III活性。结论：改进后的比色法具有较高的空白率和较高的抗霉素A敏感活性，适用于测定肝、心、骨骼肌和脑组织线粒体中的复合体III。低浓度的牛血清白蛋白和廉价洗涤剂吐温-20的使用使这种改进的方法在临床实验室环境中的使用更加稳健。(C)2008爱思唯尔B.V.保留所有权利。
Background: The determination of the activity of complex III in tissue samples provides critical evidence in the diagnosis of mitochondrial disorders and diseases associated with mitochondrial dysfunction. However, great variations have been seen in the literature due to the use of different assays.Methods: Reaction conditions of an improved spectrophotometric method exhibiting higher specificity for complex III activity than the methods currently used, were studied.Results: Optimum conditions, using bovine serum albumin at 0.01% and Tween-20 at 0.05%, were defined. The present method possesses more antimycin A-sensitive complex III activity, compared to previous methods. Thus, this improved method is sensitive and suitable for assaying complex III in both crude tissue homogenate and isolated mitochondria of liver, heart, skeletal muscle and brain.Conclusions: This spectrophotometric assay is sensitive, and specific for complex III activity because of the negligible blank rate and high antimycin A-sensitive activity. The low concentration of bovine serum albumin, and the use of inexpensive detergent Tween-20 make this improved method more robust for its use in a clinical laboratory setting. (C) 2008 Elsevier B.V. All rights reserved.