The complexity of the secreted NPA and FAR lipid-binding protein families of Haemonchus contortus revealed by an iterative proteomics-bioinformatics approach
The complexity of the secreted NPA and FAR lipid-binding protein families of Haemonchus contortus revealed by an iterative proteomics-bioinformatics approach
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DOI：
10.1016/j.molbiopara.2009.07.001
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发表时间：
2009-11-01
影响因子：
1.5
通讯作者：
Wijffels, Gene
中科院分区：
文献类型：
--
作者：
Kuang, Lisha;Colgrave, Michelle L.;Wijffels, Gene
线虫寄生虫分泌两类不同的小分子脂类结合蛋白，即线虫多蛋白过敏原/抗原(NPAs)和脂肪酸和视黄醇结合蛋白(FAR)。到目前为止，还没有证据表明这两个蛋白家族在螺旋血吸虫中表达或分泌。在本研究中，我们应用蛋白质组学和生物信息学工具，在蛋白质和基因水平上揭示了这些蛋白质在螺旋体成虫排泄/分泌产物(ESP)中的表达及其复杂性。根据标准数据库对ESP组分的质谱图进行初步检查，得到9个与Osteragia osteragi NPA和FAR序列相对应的多肽。用osteragi和Canowhabditis elegans同源物搜索缠绕线虫EST和基因组重叠群数据库，检索到编码扭转线虫NPA和FAR蛋白的不同序列。然后将H.Conortus序列整合到定制的数据库中，并且对质谱图的新搜索实现了对预测的H.Conortus NPA的10倍的覆盖。最终的质谱学分析结果表明，对扭曲线虫NPA的覆盖率为49%-60%，对扭曲线虫的覆盖率为7%-47%。此外，通过将基因组序列数据与预测的蛋白质序列进行组合，揭示了编码基因结构的多样性。我们预测在轮纹夜蛾中至少有1个HC-NPA基因和6个HC-Far基因，并且HC-Far-1到-6的生活期基因表达显示出这些基因各自独特的转录模式。皇冠版权所有(C)2009由爱思唯尔出版。保留所有权利。
Two different classes of small nematode specific lipid-binding proteins, the nematode polyprotein allergens/antigens (NPAs) and the fatty acid- and retinol-binding (FAR) proteins, are secreted by helminth parasites. Until now, there was no evidence of the expression or secretion of these two families of proteins in Haemonchus contortus. In this study, we applied proteomic and bioinformatic tools in an iterative manner to reveal the expression and complexity of these proteins in the excretory/secretory products (ESP) of adult H. contortus at the protein and gene levels. initial examination of the mass spectra of ESP fractions against standard databases returned nine peptides mapping to Ostertagia ostertagi NPA and FAR sequences. Searches of the H. contortus EST and genomic contig databases with the O. ostertagi and Caenorhabditis elegans homologues retrieved diverse sequences encoding H. contortus NPA and FAR proteins. H. contortus sequences were then integrated into a customized database and a new search of the mass spectra achieved a 10-fold improvement in coverage of the predicted H. contortus NPAs. The final analyses of the mass spectra achieved 49-60% coverage of H. contortus NPAs and 7-47% coverage of H. contortus FARs. Moreover, the diversity in structures of the encoding genes was revealed by assembling the genomic sequence data with predicted protein sequences confirmed by the peptide evidence. We predict there are at least one Hc-NPA gene and six Hc-FAR genes in H. contortus, and life stage gene expression of Hc-FAR-1 to -6 revealed unique transcription patterns for each of these genes. Crown Copyright (C) 2009 Published by Elsevier B.V. All rights reserved.