Schneider André, Bouzaidi-Tiali Nabile, Chanez Anne-Laure, Bulliard Laurence
Department of Biology, University of Fribourg, Switzerland.
Methods Mol Biol. 2007;372:379-87. doi: 10.1007/978-1-59745-365-3_27.
This chapter describes a luciferase-based protocol to measure adenosine triphosphate (ATP) production in isolated mitochondria of Trypanosoma brucei. The assay represents an excellent method to characterize the functionality of isolated mitochondria. Comparing the ATP production induced by substrates for oxidative phosphorylation to the one induced by substrates for substrate-level phosphorylation allows conclusions regarding the integrity of the outer and inner mitochondrial membranes. Furthermore, the assay is a valuable tool for characterization of RNA interference cell lines suspected to affect mitochondrial functions.
本章介绍了一种基于荧光素酶的方法,用于测量布氏锥虫分离线粒体中的三磷酸腺苷(ATP)生成。该测定法是表征分离线粒体功能的一种出色方法。将氧化磷酸化底物诱导的ATP生成与底物水平磷酸化底物诱导的ATP生成进行比较,可得出有关线粒体外膜和内膜完整性的结论。此外,该测定法是表征怀疑影响线粒体功能的RNA干扰细胞系的宝贵工具。