Parasite Disease Group, IBMC - Instituto de Biologia Molecular e Celular, Universidade do Porto, Portugal.
J Proteomics. 2013 Jun 12;84:106-18. doi: 10.1016/j.jprot.2013.03.012. Epub 2013 Apr 1.
The exoproteome of Leishmania infantum is composed of parasite derived proteins present in the extracellular environment. Although the exoproteome might have a significant role in the precocious steps of infection little is known concerning its composition. We developed an approach enabling the in vitro recovery of the exoproteome from logarithmic and stationary L. infantum promastigotes. The recovered exoproteomes were further separated into two fractions, vesicles and vesicle depleted exoproteome, evaluating the fraction protein profile. Although the most abundant protein in all fractions was GP63, the protein composition of the separated fractions was distinct reflecting the origin of the fraction and the metabolic state of the parasites. The vesicle-derived exoproteome recovered from logarithmic parasites was significantly enriched in ribosomal proteins, indicating a potential role for these vesicles in protein turnover. Also, a stage specific enrichment of vesicles with properties related to apoptotic vesicles was observed in stationary phase parasites and evidence was obtained that the release of vesicles was increased in response to a death stimuli. This report on the exoproteome obtained from in vitro promastigote cultures provides new perspectives on Leishmania biology with the possibility of vesicles playing a major role in protein turnover and also in cell death.
The first systematic insight into Leishmania exoproteome composition and the impact of the selected recovery approach.
利什曼原虫的外蛋白质组由存在于细胞外环境中的寄生虫衍生蛋白组成。尽管外蛋白质组可能在感染的早期阶段发挥重要作用,但关于其组成的了解甚少。我们开发了一种方法,能够从对数和静止期利什曼原虫前鞭毛体中体外回收外蛋白质组。回收的外蛋白质组进一步分为囊泡和囊泡耗尽的外蛋白质组两个部分,评估了部分蛋白质谱。虽然所有部分中最丰富的蛋白质都是 GP63,但分离部分的蛋白质组成是不同的,反映了部分的起源和寄生虫的代谢状态。从对数期寄生虫中回收的囊泡衍生的外蛋白质组富含核糖体蛋白,表明这些囊泡在蛋白质周转中可能发挥作用。此外,在静止期寄生虫中观察到具有与凋亡囊泡相关特性的囊泡的阶段特异性富集,并获得了证据表明,囊泡的释放增加是对死亡刺激的反应。本报告关于从体外前鞭毛体培养物中获得的外蛋白质组,为利什曼生物学提供了新的视角,可能是囊泡在蛋白质周转中发挥主要作用,也在细胞死亡中发挥主要作用。
首次系统地了解利什曼原虫外蛋白质组的组成以及所选回收方法的影响。