多头带绦虫成虫和幼虫的iTRAQ蛋白质组学分析

iTRAQ proteomics analysis of Taenia multiceps adults and larvae

目的分析多头带绦虫成虫和幼虫样本中的差异蛋白定量结果,为筛选调节寄生虫发育和生长的特定蛋白奠定基础。方法提取多头带绦虫成虫与脑多头蚴原头节蛋白质样本进行SDS-PAGE电泳分析后,采用同位素标记相对与绝对定量技术(iTRAQ)技术和液相串联质谱(LC-MS/MS)分析,当两种样品蛋白的差异倍数达到1.5倍以上,且经检验有统计学意义时,视为差异蛋白。结果本研究共鉴定了684个蛋白,其中130个差异表达蛋白,相对于幼虫,成虫上调蛋白69个,分别为副肌球蛋白,果糖-1,6-二磷酸酶,钙结合蛋白,六钩蚴蛋白Tso22c,热休克蛋白70,8 kDa糖蛋白,肌球蛋白轻链,钙调蛋白等;成虫下调蛋白61个,分别是抗原cC1,钙蛋白酶,膜联蛋白B3,皮层蛋白,糖脂转移蛋白,微管相关蛋白,乳酸脱氢酶B,xpa-结合蛋白等。通过实时定量PCR验证了10个差异明显的蛋白。另外,差异蛋白路径注释中约30%的蛋白参与了代谢途径和次生代谢物的生物合成。结论本研究为多头带绦虫发育、生活史及寄生虫-宿主的互作机制研究提供了基线资料。

This study investigated the differential protein expression profiles of the developmental stages of Taenia multiceps,to provide a basis for the identification of specific proteins regulating the parasite’s development and growth.Extracted proteins from Coenurus cerebralis protoscolex and adult stage were used for SDS-PAGE analysis.Protein expression profiles of T.multiceps were analyzed with iTRAQ and LC-MS/MS,and compared between adults and larvae.Comparative proteomics was performed to discover differentially expressed proteins between adults and larvae,on the basis of≥1.5-fold change in expression and P value<0.05.A total of 684 proteins were detected,and 130 differentially expressed proteins(DEPs)were defined;69 proteins were up-regulated in adults compared with larvae,including paramyosin,fructose-1,6-bisphosphataseI,calumenin,oncosphere protein Tso22c,heat shock protein 70,8 kDa glycoprotein,myosin light chain,and calmodulin;61 proteins were down-regulated in adults compared with larvae,including annexin B3,antigen CC1,tegumental protein,calpain,lactate dehydrogenase B,glycolipid transfer protein,and xpa-binding protein 1.Real-time quantitative PCR confirmed that the levels of ten selected DEPs were almost consistent with those determined through LC-MS/MS analysis.In addition,approximately 30%of the proteins in the differential protein pathway annotation are involved in the biosynthesis of metabolic pathway components and secondary metabolites.The present study provides baseline information on the development,life cycle,and host interactions of T.multiceps.