Calvaruso Maria Antonietta, Smeitink Jan, Nijtmans Leo
Nijmegen Center for Mitochondrial Disorders at the Department of Pediatrics, Laboratory of Pediatrics and Neurology, Radboud University Nijmegen Medical Centre, P.O. Box 9101, 6500 HB Nijmegen, The Netherlands.
Methods. 2008 Dec;46(4):281-7. doi: 10.1016/j.ymeth.2008.09.023. Epub 2008 Oct 21.
Defects in mitochondrial oxidative phosphorylation (OXPHOS) are a frequent cause of severe inherited metabolic disorders and also contribute to aging. The OXPHOS system constitutes five multi-subunit complexes embedded in the mitochondrial inner membrane. Correct function of this system requires proper assembly of the approximately 80 proteins in the complexes, as well as numerous assembly factors. Blue native electrophoresis has become a crucial tool to investigate OXPHOS-related defects in mitochondrial disease patients. In addition, OXPHOS-assembly profiles can be obtained by two dimensional blue native/SDS gel electrophoresis, which provides additional information for identifying disease-causing mutations and insight in the role of specific proteins in the biogenesis of the OXPHOS system. Here we provide a practical guide on how to set-up the basic technique to study OXPHOS defects in patient-derived cells and tissues.
线粒体氧化磷酸化（OXPHOS）缺陷是严重遗传性代谢紊乱的常见原因，也与衰老有关。氧化磷酸化系统由嵌入线粒体内膜的五个多亚基复合物组成。该系统的正常功能需要复合物中约80种蛋白质以及众多组装因子的正确组装。蓝色天然电泳已成为研究线粒体疾病患者氧化磷酸化相关缺陷的关键工具。此外，通过二维蓝色天然/SDS凝胶电泳可以获得氧化磷酸化组装图谱，这为识别致病突变以及深入了解特定蛋白质在氧化磷酸化系统生物发生中的作用提供了额外信息。在这里，我们提供了一份实用指南，介绍如何建立在患者来源的细胞和组织中研究氧化磷酸化缺陷的基本技术。
