Proteomics Research Center, Faculty of Paramedical Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical. Sciences, Tehran, Iran.
Microb Pathog. 2020 Dec;149:104557. doi: 10.1016/j.micpath.2020.104557. Epub 2020 Oct 2.
Leishmaniasis is an infectious disease caused by Leishmania that widespread in 98 countries. The differentiation of Leishmania (L) from procyclic to metacyclic promastigote has occurred along with morphological and biochemical changes in proteome scale. We aim here to identify the proteomes of two successive developmental forms (procyclic and metacyclic promastigotes) from Leishmania major isolates using SWATH-MS quantitative proteomics technique. Isolated proteins from procyclic and metacyclic lysate were digested, fractionated and subjected to SWATH-MS. Proteins significantly different in abundance were analyzed using gene ontology (GO) and protein-protein interaction network (PPIN). Our study showed that 52 proteins were changed in abundance between the two consecutive developmental stages. Differentially expressed proteins were classified into nine classes by GO analysis. Significant modulations in translation, antioxidant and stress-related defenses, energy metabolism, structural and motility-related proteins were detected between procyclic and metacyclic stages. We found that elongation factor-2 and various structural constituents of ribosome were down-regulated during metacyclogenesis, while motility related proteins including ADP-ribosylation factor-3, paraflegellar rod protein-2C and tubulin alpha-chain were up regulated. According to network analysis, ENOL has been introduced as main hub-bottleneck protein and EF-1b, Hsp60 and GDH have been determined as seed proteins. Our results show that significant proteins in abundance are crucial features of metacyclogenesis in L. major. The protein function analysis illustrated that synthetic pathway involved proteins were down-regulated in metacyclic, which is the main feature of this stage of parasite growth cycle, while up-regulation of motility and energy metabolism related proteins is consistent with infective feature of metacyclic stage. Based on our results, we suppose that differentially expressed proteins possibly play a critical role in L. major differentiation. In addition, our finding demonstrated the possibility of SWATH-MS as viable technique to faster detect new stage-specific proteins in Leishmania and further studies are required for the validation of the results.
利什曼病是一种由利什曼原虫引起的传染病，广泛分布于 98 个国家。利什曼原虫从前cyclic 到 metacyclic 前鞭毛体的分化伴随着蛋白质组规模的形态和生化变化。我们旨在使用 SWATH-MS 定量蛋白质组学技术鉴定来自利什曼主要分离株的两种连续发育形式（前cyclic 和 metacyclic 前鞭毛体）的蛋白质组。从前cyclic 和 metacyclic 裂解物中分离出的蛋白质进行消化、分级，并进行 SWATH-MS 分析。使用基因本体论 (GO) 和蛋白质-蛋白质相互作用网络 (PPIN) 分析丰度差异显著的蛋白质。我们的研究表明，在两个连续的发育阶段之间，有 52 种蛋白质的丰度发生了变化。通过 GO 分析，差异表达蛋白被分为九类。在前cyclic 和 metacyclic 阶段之间检测到翻译、抗氧化和应激防御、能量代谢、结构和运动相关蛋白的显著调节。我们发现，延伸因子-2 和核糖体的各种结构成分在前鞭毛体发生过程中下调，而运动相关蛋白，包括 ADP-ribosylation factor-3、paraflagellar rod protein-2C 和微管 alpha 链上调。根据网络分析，ENOL 被引入为主要枢纽-瓶颈蛋白，EF-1b、Hsp60 和 GDH 被确定为种子蛋白。我们的结果表明，丰度显著的蛋白质是 L. major 前鞭毛体发生的重要特征。蛋白质功能分析表明，合成途径涉及的蛋白质在前鞭毛体中下调，这是寄生虫生长周期这一阶段的主要特征，而运动和能量代谢相关蛋白的上调与前鞭毛体阶段的感染特征一致。基于我们的结果，我们假设差异表达蛋白可能在前鞭毛体分化中发挥关键作用。此外，我们的发现表明 SWATH-MS 作为一种可行的技术来更快地检测利什曼虫中的新阶段特异性蛋白是可能的，需要进一步的研究来验证结果。
