Mojtahedi Zahra, Clos Joachim, Kamali-Sarvestani Eskandar
Department of Immunology, Shiraz University of Medical Sciences, Shiraz Medical School, P.O. Box 71345-1798, Shiraz 734-8553, Iran.
Exp Parasitol. 2008 Jul;119(3):422-9. doi: 10.1016/j.exppara.2008.04.008. Epub 2008 Apr 14.
The differentiation from procyclic to metacyclic promastigotes (metacyclogenesis) has been correlated with an increased infectivity in a number of Leishmania species. We compared the proteomes of procyclic and metacyclic promastigotes of L. major. Lysates from either life cycle stage were resolved by 2D-PAGE, followed by Coomassie brilliant blue staining. Spots were analyzed by MALDI-TOF MS. 25 protein spots were found to be differentially expressed during metacyclogenesis. We found that proteins involved in protein synthesis were less abundant in metacyclic promastigotes, while proteins involved in motility, including paraflagellar rod protein 1D, alpha-tubulin and beta-tubulin were more abundant. Also, two mitochondrial enzymes (succinyl-CoA synthetase beta subunit and cytochrome c oxidase subunit IV) were differentially expressed in both life cycle stages. Down-regulation of proteins related to synthetic pathway in metacyclic promastigotes is consistent with the arrested growth in this life cycle stage, while up-regulation of proteins related to motility in metacyclic promastigotes is in agreement with the high motility observed in this stage.
在许多利什曼原虫物种中,前鞭毛体向循环后期前鞭毛体的分化(循环后期发育)与感染力增加有关。我们比较了硕大利什曼原虫前鞭毛体和循环后期前鞭毛体的蛋白质组。来自任一生命周期阶段的裂解物通过二维聚丙烯酰胺凝胶电泳进行分离,随后进行考马斯亮蓝染色。通过基质辅助激光解吸电离飞行时间质谱对斑点进行分析。发现在循环后期发育过程中有25个蛋白斑点存在差异表达。我们发现,参与蛋白质合成的蛋白质在循环后期前鞭毛体中含量较少,而参与运动的蛋白质,包括副鞭毛杆蛋白1D、α-微管蛋白和β-微管蛋白含量较多。此外,两种线粒体酶(琥珀酰辅酶A合成酶β亚基和细胞色素c氧化酶亚基IV)在两个生命周期阶段均有差异表达。循环后期前鞭毛体中与合成途径相关的蛋白质下调与该生命周期阶段生长停滞一致,而循环后期前鞭毛体中与运动相关的蛋白质上调与该阶段观察到的高运动性一致。