紫外线照射前后日本血吸虫尾蚴可溶性虫体蛋白组分的比较

Schistosoma japonicum: Proteomic Analysis of Ultraviolet-Attenuated Cercariae and the Normal Cercariae

目的利用蛋白质组学技术分离、鉴定日本血吸虫正常尾蚴及紫外线致弱尾蚴虫体间差异表达蛋白。方法分别收集与制备血吸虫正常尾蚴和致弱尾蚴虫体总蛋白,经固相pH梯度双向凝胶电泳分离。凝胶银染并利用ImageMaster2DSoftware5.0凝胶图像分析软件进行比较分析,选取差异表达蛋白点经基质辅助激光解析离子飞行时间质谱仪进行鉴定。结果二维凝胶电泳图像分析结果显示:正常尾蚴和致弱尾蚴各分离出至少1277、1173个清晰、独立的蛋白点,蛋白点匹配率为72.7%,大多数蛋白点相对分子量处于22000～95000范围内,理论等电点为5～8。尾蚴经紫外线致弱前后共发现18个差异表达蛋白,其中5个蛋白于致弱尾蚴中表达消失,12个表达下调,1个表达增强,未见新增表达蛋白。其中13个差异表达蛋白经MALDI-TOF-MS鉴定分析,获悉其肽指纹图谱、理论等电点、分子量及表达水平变化等相关信息。结论利用二维电泳、MALDI-TOF-MS等蛋白质组学技术,共鉴定出13个致弱尾蚴差异表达蛋白,为进一步阐明致弱尾蚴诱导宿主产生高免疫保护力的分子机制提供了实验基础。

Objective To examine the protein expression profiles of normal and ultraviolet light （UV） attenuated Schistosonva japonicum eercafiae. Methods Total proteins were prepared from normal and UV attenuated cercariae. After two-dimensional （2-D） gel electrophoresis, distinct protein spots from 2-D gels were isolated and analyzed by MALDI-TOF MS. Results A total of 1 277 （from normal cereariae） and 1 173 （from UV attenuated cercariae） protein spots were detected in the 2-D gels. Most of the proteins had a molecular weight of 22 000 to 95 000, and a theoretical pI （isoeleetrie point） value of 5 to 8. Seventy-two percent of the protein spots from UV attenulated group was found to match with the normal untreated group. Five proteins were turned off in expression and 13 proteins were differentially expressed after irradiation. Thirteen out of the 18 distinct protein spots were selected and identified by MALDI-TOF MS and their pI, molecular weight, and expression levels were obtained. ConclusionEighteen proteins were differentially expressed after UV irradiation. Thirteen differentially expressed proteins were identified by 2-D electrophoresis and MALDI-TOF-MS. Further research on these differentially expressed proteins might enhance our understanding on the protection conferred by UV attenuated cercariae.