GPIomics: global analysis of glycosylphosphatidylinositol-anchored molecules of Trypanosoma cruzi
GPIomics: global analysis of glycosylphosphatidylinositol-anchored molecules of Trypanosoma cruzi
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DOI：
10.1038/msb.2009.13
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发表时间：
2009-04-01
影响因子：
9.9
通讯作者：
Almeida, Igor C.
中科院分区：
文献类型：
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作者：
Nakayasu, Ernesto S.;Yashunsky, Dmitry V.;Almeida, Igor C.
糖基磷脂酰肌醇（GPI）锚定是一种常见的真核表面蛋白的翻译后修饰。在这里，我们开发了一种快速、简单、高灵敏度（高原子摩尔-低飞摩尔范围）的方法，该方法使用液相色谱-串联质谱（LC-MSn）对真核生物克氏锥虫（恰加斯病的病原）的gpi锚定分子（即GPIome）进行了首次大规模分析。我们的基因组预测分析显示，大约12%的克氏锥虫基因可能编码gpi锚定蛋白。利用LC-MSn对克氏绦虫居虫期的GPIome进行分析，鉴定出90种GPIome，其中79种为新种。此外，我们还确定了克氏t小黏液样基因（TcSMUG S）家族编码的黏液蛋白是表皮毛囊细胞表面表达的主要gpi锚定蛋白。TcSMUG S粘蛋白成熟序列较短（56-85个氨基酸），o -糖基化程度高，含有较少的蛋白水解位点，因此对昆虫媒介中肠蛋白酶不太敏感。我们建议我们的方法可以用于其他低等和高等真核生物的高通量gpiomics分析。分子系统生物学2009年4月7日；doi: 10.1038 / msb.2009.13
Glycosylphosphatidylinositol (GPI) anchoring is a common, relevant posttranslational modification of eukaryotic surface proteins. Here, we developed a fast, simple, and highly sensitive (high attomole-low femtomole range) method that uses liquid chromatography-tandem mass spectrometry (LC-MSn) for the first large-scale analysis of GPI-anchored molecules (i.e., the GPIome) of a eukaryote, Trypanosoma cruzi, the etiologic agent of Chagas disease. Our genome-wise prediction analysis revealed that approximately 12% of T. cruzi genes possibly encode GPI-anchored proteins. By analyzing the GPIome of T. cruzi insect-dwelling epimastigote stage using LC-MSn, we identified 90 GPI species, of which 79 were novel. Moreover, we determined that mucins coded by the T. cruzi small mucin-like gene (TcSMUG S) family are the major GPI-anchored proteins expressed on the epimastigote cell surface. TcSMUG S mucin mature sequences are short (56-85 amino acids) and highly O-glycosylated, and contain few proteolytic sites, therefore, less likely susceptible to proteases of the midgut of the insect vector. We propose that our approach could be used for the high throughput GPIomic analysis of other lower and higher eukaryotes. Molecular Systems Biology 7 April 2009; doi:10.1038/msb.2009.13