不同种株利什曼原虫的比较蛋白质组学分析

Comparative proteomics analysis of different Leishmania spp.

目的观察不同种株利什曼原虫前鞭毛体蛋白质表达状况。方法制备杜氏利什曼原虫四川SC6株、杜氏利什曼原虫四川SC10株和硕大利什曼原虫5ASKH株前鞭毛体总蛋白,以pH范围3-10的预制胶条进行双向电泳(2-D),考马斯亮蓝染色,PDQust软件分析凝胶,主要差异蛋白点用电喷雾质谱法进行鉴定。结果等量的杜氏利什曼原虫四川SC6株、杜氏利什曼原虫四川SC10株和硕大利什曼原虫5ASKH株前鞭毛体总蛋白均获近700个蛋白点,不同种株利什曼原虫前鞭毛体蛋白质2-D图谱中,14蛋白点呈恒定差异表达,从中鉴定出10个功能明确的蛋白质,分别具有下列生物功能:糖代谢与磷脂合成(烯醇酶、变旋酶、NADP依耐乙醇脱氢酶、乙醇胺磷酸胞苷酸转移酶),压力反应(细胞内过氧化物酶、锥虫还原蛋白过氧化物酶),细胞膜/细胞骨架(α-微管蛋白、β-微管蛋白),核酸代谢(琥珀酰辅酶A连接酶(GDP形成)、内源性RNA酶L-PSP(pb5)),细胞周期与增殖(延伸因子2)。结论不同种株利什曼原虫前鞭毛体蛋白质的表达存在不同,为理解不同种株利什曼原虫的毒力、免疫原性和代谢特征提供了新的视角。

Leishmaniasis is a zoonotic disease with diverse clinical manifestations and caused by different species and strains of the Leishmania genus. In the present study, the protein variability in the promastigotes of different Leishmania spp. , such as Leishmanis major strain 5ASKH. Leishmania donovani strain SC6 and strain SC10 was identified through proteomies analysis in which resolution of the soluble proteomes of axenic promastigotes was investigated by two dimensional eleetrophoresis （2DE） in the broad pH range （3 10）. It was demonstrated that equivalent numbers of protein spots could be found in each strain of Leishmania species and a total of near 700 protein spots were visualized by using Coomassie Blue staining. In the proteomes of promastigotes of different strains, 10 defined proteins identified from 14 protein spots differentially expressed in distinct Leishmania spp. were demonstrated and they fall in several functional classes, i.e. carbobydrate/energy metabolism and phosphorylation （enolase, aldose 1-epimerase, NADP.dependent alcohol dehydrogenase, ethanolamine-phosphate eytidylyl- transferase ）, stress response （eytosolie peroxiredoxin, tryparedoxin peroxidase ）, cell membrane/eytoskeleton （α-tubulin, β- tubulin. ）, nueleotide acid metabolism Esuccinyl CoA ligase（GDP-forming） β-chain, endoribonuelease L-PSP （pb5）], cell cycle and proliferation （elongation factor 2 ）. It is evident that different protein expression profile of the promastigotes exists in different Leishmania spp. and this finding may bring into a new insight in our understanding on the virulence, immunogenieity and metabolism characteristics of Leishrnania.