Analysis of Mitochondrial Respiratory Chain Supercomplexes Using Blue Native Polyacrylamide Gel Electrophoresis (BN‐PAGE)

Q1 Agricultural and Biological Sciences

Current protocols in mouse biology Pub Date : 2016-03-01 DOI:10.1002/9780470942390.mo150182

Pooja Jha, Xu Wang, J. Auwerx

引用次数: 176

Abstract

Mitochondria are cellular organelles that harvest energy in the form of ATP through a process termed oxidative phosphorylation (OXPHOS), which occurs via the protein complexes of the electron transport chain (ETC). In recent years it has become unequivocally clear that mitochondrial complexes of the ETC are not static entities in the inner mitochondrial membrane. These complexes are dynamic and in mammals they aggregate in different stoichiometric combinations to form supercomplexes (SCs) or respirasomes. It has been proposed that the net respiration is more efficient via SCs than via isolated complexes. However, it still needs to be determined whether the activity of a particular SC is associated with a disease etiology. Here we describe a simplified method to visualize and assess in‐gel activity of SCs and the individual complexes with good resolution using blue native polyacrylamide gel electrophoresis (BN‐PAGE). © 2016 by John Wiley & Sons, Inc.

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蓝色天然聚丙烯酰胺凝胶电泳(BN‐PAGE)分析线粒体呼吸链超配合物

线粒体是一种细胞器，通过称为氧化磷酸化(OXPHOS)的过程以ATP的形式收集能量，该过程通过电子传递链(ETC)的蛋白质复合物发生。近年来，已经明确地表明，ETC的线粒体复合体不是线粒体膜内的静态实体。这些复合物是动态的，在哺乳动物中，它们以不同的化学计量组合聚集形成超复合物(SCs)或呼吸小体。有人提出，净呼吸通过SCs比通过孤立的配合物更有效。然而，仍然需要确定特定SC的活性是否与疾病病因有关。在这里，我们描述了一种简化的方法，使用蓝色天然聚丙烯酰胺凝胶电泳(BN - PAGE)，以良好的分辨率可视化和评估SCs和单个复合物的凝胶活性。©2016 by John Wiley & Sons, Inc。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Current protocols in mouse biology Agricultural and Biological Sciences-Animal Science and Zoology

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期刊介绍： Sound and reproducible laboratory methods are the foundation of scientific discovery. Yet, all too often, nuances that are critical for an experiment''s success are not captured in the primary literature but exist only as part of a lab''s oral tradition. The aim of Current Protocols in Mouse Biology is to provide the clearest, most detailed and reliable step-by-step instructions for protocols involving the use of mice in biomedical research. Written by experts in the field and extensively edited to our exacting standards, the protocols include all of the information necessary to complete an experiment in the laboratory—introduction, materials lists with supplier information, detailed step-by-step procedures with helpful annotations, recipes for reagents and solutions, illustrative figures and information-packed tables. Each article also provides invaluable discussions of background information, applications of the methods, important assumptions, key parameters, time considerations, and tips to help avoid common pitfalls and troubleshoot experiments. Furthermore, Current Protocols in Mouse Biology content is thoughtfully organized by topic for optimal usage and to maximize contextual knowledge. Quarterly issues allow Current Protocols to constantly evolve to keep pace with the newest discoveries and developments. Current Protocols in Mouse Biology brings together resources in mouse biology and genetics and provides a mouse protocol resource that covers all aspects of mouse biology. Current Protocols in Mouse Biology also permits optimization of mouse model usage, which is significantly impacted by both cost and ethical constraints. Optimal and standardized mouse protocols ultimately reduce experimental variability and reduce the number of animals used in mouse experiments.