Monitoring the Uptake of Glycosphingolipids In Plasmodium falciparum-Infected Erythrocytes Using Both Fluorescence Microscopy and Capillary Electrophoresis with Laser-Induced Fluorescence Detection
Monitoring the Uptake of Glycosphingolipids In Plasmodium falciparum-Infected Erythrocytes Using Both Fluorescence Microscopy and Capillary Electrophoresis with Laser-Induced Fluorescence Detection
复制标题
DOI：
10.1021/ac1021776
复制
发表时间：
2010-12-01
影响因子：
7.4
通讯作者：
Dovichi, Norman J.
中科院分区：
文献类型：
--
作者：
Essaka, David C.;White, John;Dovichi, Norman J.
恶性疟原虫对鞘糖脂的代谢在疾病的发展过程中起着重要作用。本文报道了一种监测感染红细胞摄取鞘糖脂的新的、高灵敏度的方法。以四甲基罗丹明标记的鞘糖脂(GM1-TMR)为底物，在荧光显微镜下观察摄取。红细胞用15%的皂苷溶液裂解，用磷酸盐缓冲盐水洗涤，释放完整的寄生虫。寄生虫进一步裂解，所得匀浆用毛细管电泳法和激光诱导荧光检测分析。在1%寄生虫血症时，感染红细胞的裂解液产生的标准偏差比未感染的红细胞大20个标准差。这表明可以用这种技术研究相对较低的感染水平
The metabolism of glycosphingolipids by the malaria-causing parasite Plasmodium falciparum plays an important role in the progression of the disease We report a new and highly sensitive method to monitor the uptake of glycosphingolipids in infected red blood cells (iRBCs) A tetramethylrhodamine-labeled glycosphingolipid (GM1 TMR) was used as a substrate Uptake was demonstrated by fluorescence microscopy The IRBCs were lysed with a 15% solution of saponin and washed with phosphate buffered saline to release intact parasites The parasites were further lysed and the resulting homogenates were analyzed by capillary electrophoresis with laser-induced fluorescence detection The lysate from erythrocytes infected at 1% parasitemia generated a signal 20 standard deviations larger than uninfected erythrocytes, which suggests that relatively low infection levels can be studied with this technique