Two-dimensional electrophoresis and mass spectrometry for the identification of species-specific Trichinella antigens
Two-dimensional electrophoresis and mass spectrometry for the identification of species-specific Trichinella antigens
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DOI：
10.1016/j.vetpar.2005.05.018
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发表时间：
2005-09-05
影响因子：
2.6
通讯作者：
Bolás-Fernández, F
中科院分区：
文献类型：
--
作者：
Dea-Ayuela, MA;Bolás-Fernández, F
利用蛋白质组学方法对密切相关的旋毛虫基因型布氏旋毛虫T3和旋毛虫T8进行精细抗原表征。用213-PAGE分析了两个分离物的肌幼虫L1（LCE）粗提物。在两种基因型中，超过500个蛋白质点可重复地分离。这些分离的蛋白质与IgG 1和IgG 3的同源和异源的超免疫血清中提出的BALB/c小鼠进行了鉴定。在布氏锥虫和旋毛虫T8图谱中，分别在50-60 kDa和pH 5.5-6.5处迁移的一组20和15个斑点与来自异源血清的IgGI反应，而一组具有相似迁移模式的次要斑点则不反应。IgG 3的交叉反应性较低。这些抗原的MALDI-TOF和MALDI-TOF/TOF分析确定了预测的烯醇化酶，蛋白质P49和预测的肌动蛋白之间的交叉反应蛋白和两个假设的肌动蛋白之间的非交叉反应蛋白。(c)2005 Elsevier B. V.保留所有权利。
A proteomic approach was utilized for fine antigenic characterization of the closely related Trichinella genotypes Trichinella britovi T3 and Trichinella T8. Crude extract of muscle larvae L1 (LCE) from both isolates were analyzed by 213-PAGE. Over 500 protein spots were reproducibly separated in both genotypes. These separated proteins were identified in Western blot with IgG1 and IgG3 from homologous and heterologous hyperimmune sera raised in BALB/c mice. A group of 20 and 15 spots migrating at 50-60 kDa and pH 5.5-6.5 in T britovi and Trichinella T8 maps, respectively, reacted with the IgGI from heterologous sera whereas a group of minor spots of similar migration patterns did not. Low cross-reactivity occurred for IgG3. MALDI-TOF and MALDI-TOF/TOF analysis of these antigens identified a predicted enolase, the protein P49 and a predicted actin among the cross-reactive proteins and two hypothetical actins among the non cross-reactive proteins. (c) 2005 Elsevier B.V. All rights reserved.