Mishra Suman, Mishra Rajnikant
Biochemistry and Molecular Biology Lab, Department of Zoology, Banaras Hindu University, Varanasi 221005, India.
Int J Proteomics. 2015;2015:647408. doi: 10.1155/2015/647408. Epub 2015 Dec 10.
Potassium chloride (KCl) has been commonly used in homogenization buffer and procedures of protein extraction. It is known to facilitate release of membrane-associated molecules but the higher concentration of KCl may affect the integrity of mitochondria by breaching the electrostatic force between the lipids and proteins. Therefore, it has been intended to explore the effect of KCl on mitochondrial proteome. The mitochondria were isolated from the mice liver and sub-fractionated into mitochondrial matrix and outer mitochondrial membrane fraction. The fractions were analysed by denaturing polyacrylamide gel electrophoresis (PAGE) and 2D-PAGE. The analysis of ultrastructure and protein profiles by MALDI-MS and data-mining reveals KCl-associated alterations in the integrity of mitochondria and its proteome. The mitochondrial membrane, cristae, and the matrix proteins appear altered under the influence of KCl.
氯化钾(KCl)常用于匀浆缓冲液和蛋白质提取过程。已知它有助于膜相关分子的释放,但较高浓度的KCl可能会破坏脂质与蛋白质之间的静电力,从而影响线粒体的完整性。因此,旨在探究KCl对线粒体蛋白质组的影响。从小鼠肝脏中分离出线粒体,并将其亚分级为线粒体基质和线粒体外膜部分。通过变性聚丙烯酰胺凝胶电泳(PAGE)和二维PAGE对这些部分进行分析。通过基质辅助激光解吸电离质谱(MALDI-MS)和数据挖掘对超微结构和蛋白质谱进行分析,揭示了与KCl相关的线粒体完整性及其蛋白质组的改变。在KCl的影响下,线粒体膜、嵴和基质蛋白似乎发生了变化。