Fuller Kathryn M, Arriaga Edgar A
Department of Chemistry, University of Minnesota, 207 Pleasant St SE, Minneapolis, MN, 55455, USA.
J Chromatogr B Analyt Technol Biomed Life Sci. 2004 Jul 5;806(2):151-9. doi: 10.1016/j.jchromb.2004.03.050.
The presence of electrical charges on the surface of an organelle is the source of the organelle's electrophoretic mobility. Recently, we reported that capillary electrophoresis with laser-induced fluorescence detection (CE-LIF) can be used to determine the electrophoretic mobility of individual mitochondria. Here, we describe the use of CE-LIF to monitor changes in the electrophoretic mobility distributions of: (i). mitochondria isolated from cultured NS-1 mouse hybridoma cells disrupted by nitrogen cavitation or mechanical homogenization; (ii). mitochondria isolated from rat liver and purified by gradient centrifugation before and after being frozen in liquid nitrogen; and (iii). mitochondria chemically transformed into mitoplasts. These results indicate that the organelle electrophoretic mobility observed by researchers is affected by preparation procedures and that CE-LIF is a complementary technique for monitoring the quality of mitochondrial preparations.
细胞器表面电荷的存在是其电泳迁移率的来源。最近,我们报道了采用激光诱导荧光检测的毛细管电泳(CE-LIF)可用于测定单个线粒体的电泳迁移率。在此,我们描述了使用CE-LIF监测以下物质电泳迁移率分布变化的情况:(i)从经氮空化或机械匀浆破坏的培养NS-1小鼠杂交瘤细胞中分离得到的线粒体;(ii)从大鼠肝脏中分离并经梯度离心纯化,在液氮中冷冻前后的线粒体;以及(iii)化学转化为线粒体膜间腔的线粒体。这些结果表明,研究人员观察到的细胞器电泳迁移率受制备程序的影响,并且CE-LIF是监测线粒体制备质量的一种补充技术。