Chemale Gustavo, Morphew Russell, Moxon Joseph V, Morassuti Alessandra L, Lacourse E James, Barrett John, Johnston David A, Brophy Peter M
School of Biological Sciences, University of Liverpool, Liverpool, UK.
Proteomics. 2006 Dec;6(23):6263-73. doi: 10.1002/pmic.200600499.
The parasite Fasciola hepatica causes major global disease of livestock, with increasing reports of human infection. Vaccine candidates with varying protection rates have been identified by pre-genomic approaches. As many candidates are part of protein superfamilies, sub-proteomics offers new possibilities to systematically reveal the relative importance of individual family proteins to vaccine formulations within populations. The superfamily glutathione transferase (GST) from liver fluke has phase II detoxification and housekeeping roles, and has been shown to contain protective vaccine candidates. GST were purified from cytosolic fractions of adult flukes using glutathione- and S-hexylglutathione-agarose, separated by 2-DE, and identified by MS/MS, with the support of a liver fluke EST database. All previously described F. hepatica GST isoforms were identified in 2-DE. Amongst the isoforms mapped by 2-DE, a new GST, closely related to the Sigma class enzymes is described for the first time in the liver fluke. We also describe cDNA encoding putative Omega class GST in F. hepatica.
肝片吸虫这种寄生虫会引发影响全球家畜的主要疾病，且人类感染病例的报告日益增多。通过基因组学之前的方法已经鉴定出了具有不同保护率的候选疫苗。由于许多候选疫苗是蛋白质超家族的一部分，亚蛋白质组学为系统揭示个体家族蛋白在群体疫苗配方中的相对重要性提供了新的可能性。来自肝吸虫的谷胱甘肽转移酶（GST）超家族具有II期解毒和看家功能，并且已被证明含有保护性候选疫苗。利用谷胱甘肽和S - 己基谷胱甘肽琼脂糖从成虫的胞质部分纯化GST，通过二维电泳（2-DE）分离，并在肝吸虫EST数据库的支持下通过串联质谱（MS/MS）进行鉴定。在二维电泳中鉴定出了所有先前描述的肝片吸虫GST同工型。在通过二维电泳定位的同工型中，首次在肝吸虫中描述了一种与西格玛类酶密切相关的新型GST。我们还描述了肝片吸虫中编码假定的欧米伽类GST的cDNA。
