Quantitative proteomic analysis reveals differentially expressed proteins in Leishmania major metacyclogenesis
Quantitative proteomic analysis reveals differentially expressed proteins in Leishmania major metacyclogenesis
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DOI：
10.1016/j.micpath.2020.104557
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发表时间：
2020-12-01
影响因子：
3.8
通讯作者：
Ahmadi, Nayebali
中科院分区：
文献类型：
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作者：
Amiri-Dashatan, Nasrin;Rezaei-Tavirani, Mostafa;Ahmadi, Nayebali
利什曼病是一种由利什曼原虫引起的传染病，广泛传播于98个国家。利什曼原虫（L）从原环前鞭毛体向中环前鞭毛体的分化伴随着蛋白质组尺度上的形态学和生化变化的沿着发生。我们的目的是在这里确定两个连续的发展形式（前环和后环前鞭毛体）从利什曼原虫分离株使用SWATH-MS定量蛋白质组学技术。从前环和后环裂解物中分离的蛋白质被消化，分级分离并进行SWATH-MS。使用基因本体（GO）和蛋白质-蛋白质相互作用网络（PPIN）分析丰度显著不同的蛋白质。我们的研究表明，52个蛋白质的丰度变化之间的两个连续的发展阶段。通过GO分析将差异表达蛋白分为9类。翻译，抗氧化和应激相关的防御，能量代谢，结构和运动相关的蛋白质的显着调制检测之间的前循环和后循环阶段。结果表明，在后循环发生过程中，延伸因子-2和核糖体的各种结构成分表达下调，而运动相关蛋白如ADP-核糖基化因子-3、鞭毛旁杆蛋白-2C和微管蛋白α链表达上调。通过网络分析，确定ENOL为主要的枢纽-瓶颈蛋白，EF-1b、Hsp 60和GDH为种子蛋白。我们的研究结果表明，丰富的重要蛋白质是L。少校蛋白质功能分析表明，代谢途径相关蛋白在代谢循环中表达下调，这是该阶段寄生虫生长周期的主要特征，而运动和能量代谢相关蛋白表达上调则与代谢循环阶段的感染特征相一致。根据我们的研究结果，我们推测差异表达的蛋白质可能在L.主要区别。此外，我们的研究结果表明SWATH-MS作为可行的技术来更快地检测利什曼原虫中新的阶段特异性蛋白的可能性，并且需要进一步的研究来验证结果。
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.