Martinez J, Perez-Serrano J, Bernadina W E, Rodriguez-Caabeiro F
Facultad de Farmacia, Departamento de Microbiologia y Parasitologia, Universidad de Alcala, 28871 Alcala de Henares, Madrid, Spain.
J Helminthol. 2002 Sep;76(3):217-23. doi: 10.1079/JOH2002127.
Stress response and phosphorylation of heat shock proteins (HSPs) 60, 70 and 90 were studied in Trichinella nativa, T. nelsoni, T. pseudospiralis and T. spiralis larvae at 30-min intervals following exposure to 20, 100 and 200 mM H2O2. There was a time- and dose-dependent differential survival for the infective stage larvae (L1) of these four Trichinella species. Immunoblotting analysis revealed that constitutive Hsp60 and Hsp70, but not Hsp90, from test Trichinella species are constitutively phosphorylated on serine/threonine residues as they converted to forms with increased sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) mobility by treatment with alkaline phosphatase. After exposure to H2O2, while there was a time-related occurrence of the three HSPs with decreased SDS-PAGE mobility, these HSPs were insensitive to alkaline phosphatase except in the case of exposure to 20 mM H2O2 for Hsp60 from all Trichinella species and Hsp70 from T. spiralis and T. nelsoni. The synthesis of HSPs forms with decreased SDS-PAGE mobility is a susceptibility signal because the lower concentration of peroxide (20 mM) did not cause a decrease on HSPs SDS-PAGE mobility in T. spiralis and T. nelsoni, the two more resistant selected Trichinella species.
在旋毛虫属的本地毛形线虫、纳氏旋毛虫、伪旋毛虫和旋毛形线虫的幼虫暴露于20、100和200 mM过氧化氢后，每隔30分钟研究一次应激反应以及热休克蛋白（HSPs）60、70和90的磷酸化情况。这四种旋毛虫属感染性幼虫（L1）的存活情况存在时间和剂量依赖性差异。免疫印迹分析显示，受试旋毛虫属物种的组成型Hsp60和Hsp70（而非Hsp90）在丝氨酸/苏氨酸残基上发生组成型磷酸化，因为经碱性磷酸酶处理后，它们转变为十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）迁移率增加的形式。暴露于过氧化氢后，虽然这三种热休克蛋白出现了SDS-PAGE迁移率下降的时间相关现象，但除了所有旋毛虫属物种的Hsp60以及旋毛形线虫和纳氏旋毛虫的Hsp70暴露于20 mM过氧化氢的情况外，这些热休克蛋白对碱性磷酸酶不敏感。SDS-PAGE迁移率下降的热休克蛋白形式的合成是一种易感性信号，因为较低浓度的过氧化物（20 mM）并未导致旋毛形线虫和纳氏旋毛虫（两种更具抗性的选定旋毛虫属物种）的热休克蛋白SDS-PAGE迁移率下降。
