Islam M K, Miyoshi T, Yamada M, Alim M A, Huang X, Motobu M, Tsuji N
Laboratory of Parasitic Diseases, National Institute of Animal Health, National Agricultural Research Organization, 3-1-5 Kannondai, Tsukuba, Ibaraki 305-0856, Japan.
Parasitology. 2006 Oct;133(Pt 4):497-508. doi: 10.1017/S0031182006000576.
Sodium fluoride (NaF) is an anion that has been previously shown to block the moulting process of Ascaris suum larvae. This study describes moulting and development-specific protein expression profiles of A. suum lung-stage L3 (AsLL3) following NaF exposure. AsLL3s cultured in the presence or absence of NaF were prepared for protein analysis using two-dimensional (2D) electrophoresis. NaF exposure inhibited at least 22 proteins in AsLL3 compared with moulted larvae (i.e. AsLL4). A further comparison of AsLL4 with those of pre-cultured AsLL3 and NaF-exposed AsLL3 revealed 8 stage-specifically and 4 over-expressed proteins. Immunoblot analysis revealed an inhibition by NaF of 19 immunoreactive proteins. Enzyme assay and immunochemical data showed an inhibition of the moulting-specific inorganic pyrophosphatase activity by 41% and a decreased expression in NaF-treated larvae, indicating its significance in the moulting process. A protein spot associated with NaF inhibition was isolated and identified by peptide mass spectrometry and bioinformatics approaches to be a member of 3-hydroxyacyl-CoA dehydrogenase/short-chain dehydrogenase enzyme families. These results have implications for the identification of proteins specific to the moulting process as potential chemotherapeutic targets.
氟化钠（NaF）是一种阴离子，先前已证明它能阻断猪蛔虫幼虫的蜕皮过程。本研究描述了暴露于NaF后猪蛔虫肺期L3幼虫（AsLL3）的蜕皮和发育特异性蛋白质表达谱。使用二维（2D）电泳对在有或没有NaF存在的情况下培养的AsLL3进行蛋白质分析准备。与蜕皮后的幼虫（即AsLL4）相比，NaF暴露抑制了AsLL3中至少22种蛋白质。将AsLL4与预培养的AsLL3和暴露于NaF的AsLL3进一步比较，发现了8种阶段特异性蛋白质和4种过表达蛋白质。免疫印迹分析显示NaF对19种免疫反应性蛋白质有抑制作用。酶活性测定和免疫化学数据表明，蜕皮特异性无机焦磷酸酶活性受到41%的抑制，并且在经NaF处理的幼虫中表达降低，表明其在蜕皮过程中的重要性。通过肽质量谱和生物信息学方法分离并鉴定了一个与NaF抑制相关的蛋白质斑点，它是3-羟基酰基辅酶A脱氢酶/短链脱氢酶酶家族的成员。这些结果对于鉴定蜕皮过程特异性蛋白质作为潜在的化疗靶点具有重要意义。
