杜氏利什曼原虫前鞭毛体和无鞭毛体的比较蛋白质组学分析  

Comparative Proteomic Analysis of the Promastigotes and Amastigotes of Leishmania donovani

作　　者：敬保迁[1] 邓世山[1] 张仁刚[1] 张洁[1] 

机构地区：[1]川北医学院免疫学与分子生物学研究所,南充637000

出　　处：《中国寄生虫学与寄生虫病杂志》2009年第2期102-106,共5页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：国家自然科学基金(No.30571635)~~

摘　　要：目的应用蛋白质组学技术分析杜氏利什曼原虫前鞭毛体和无鞭毛体蛋白质表达状况。方法分别提取和纯化杜氏利什曼原虫四川SC6株前鞭毛体与纯培养无鞭毛体的总蛋白,分别经pH3～10的预制胶条进行双向电泳分离,凝胶用考马斯亮蓝染色,凝胶图像以PDQuest1.0软件分析,并对主要差异表达蛋白点用电喷雾质谱法进行鉴定。结果等量的前鞭毛体与纯培养的无鞭毛体总蛋白经双向电泳分离后均可获近700个蛋白点,其中超过90%的蛋白点的分布和相对强度基本一致。与前鞭毛体比较,6个蛋白点在无鞭毛体蛋白中明显高表达,3个蛋白点低表达。6个明显高表达的蛋白点中有5个为已知功能蛋白,分别为Reiske铁硫蛋白前体、α微管蛋白、过氧化物酶1、二氢硫辛酰胺乙酰转移酶前体和甘露糖-1-磷酸瓜氨酸转移酶;3个低表达的蛋白点中有2个为已知功能蛋白,分别为热休克蛋白70和β微管蛋白。这些差异调节表达蛋白与碳水化合物/能量代谢,应激反应,细胞膜和细胞骨架形成相关。结论前鞭毛体与无鞭毛体蛋白质的表达存在差异。Objective To explore the protein profile and identify developmentally regulated proteins of the promastigotes and axenic amastigotes with comparative proteomics technique. Methods The total proteins of promastigotes and axenic amastigotes of Leishmania donovani SC6 strain were separated by two-dimensional electrophoresis（2-DE） in a broad pH range（3-10）,and the gel was stained with Coomassie blue. The images were analyzed by PDQuest 1.0 software,and the major developmentally regulated proteins were identified by electrospray mass spectrometry. Results Approximately 700 protein spots were revealed in equivalent proteins of the promastigotes and axenic amastigotes separated by 2-DE,among which more than 90% protein spots showed equivalent quantity and distribution,with 6 proteins up-regulated and 3 proteins down-regulated in axenic amastigotes compared with promastigotes. Five of the 6 up-regulated proteins were with known function,respectively ascribed as Reiske iron-sulfur protein precursor,α-tubulin,peroxidoxin 1,dihydrolipoamide acetyltransferase precursor,and mannose-1-phosphate guanyltransferase. Two of the 3 down-regulated proteins were identified as heat shock protein 70 and β-tubulin. The functions of the developmentally regulated proteins were related to the carbohydrate/energy metabolism,stress response,or formation of cell membrane/cytoskeleton. Conclusion The findings demonstrate the differences in protein expression profiles between promastigotes and amastigotes.

关 键 词：杜氏利什曼原虫 前鞭毛体 无鞭毛体 比较蛋白质组学 

分 类 号：R382.22[医药卫生—医学寄生虫学]