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病引起的传染病，在98个国家中广泛存在。利什曼原虫（L）从前环前鞭毛体向中期环前鞭毛体的分化已经发生，并且蛋白质组规模发生了形态和生化变化。我们的目的是鉴定来自利什曼原虫的两个连续发育形式（前环和后环前鞭毛体）的蛋白质组。使用SWATH-MS定量蛋白质组学技术分离菌株。消化来自前环和间环裂解物的分离蛋白，进行分级分离，然后进行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）分析了丰度显着不同的蛋白质。我们的研究表明，在两个连续的发育阶段之间，共有52种蛋白质发生了变化。通过GO分析将差异表达的蛋白质分为九类。在前循环和后循环阶段之间检测到翻译，抗氧化剂和应激相关防御，能量代谢，结构和运动相关蛋白的显着调节。我们发现，延伸因子2和核糖体的各种结构成分在细胞周期形成过程中被下调，而与运动相关的蛋白质，包括ADP-核糖基化因子3，副飞节杆蛋白2C和微管蛋白α链被上调。根据网络分析，已引入ENOL作为主要的枢纽瓶颈蛋白，并已确定EF-1b，Hsp60和GDH为种子蛋白。我们的结果表明，大量的重要蛋白质是肝细胞发生新细胞周期的关键特征。L.少校。蛋白质功能分析表明，合成途径中涉及的蛋白质在代谢循环中被下调，这是该寄生虫生长周期这一阶段的主要特征，而动力和能量代谢相关蛋白质的上调与代谢循环的感染特征相一致。根据我们的结果，我们认为差异表达的蛋白质可能在大麦芽孢杆菌的分化中起关键作用。此外，我们的发现证明了SWATH-MS作为快速检测利什曼原虫中新的阶段特异性蛋白的可行技术的可能性，并且需要进一步的研究来验证结果。