Pardo M, Ward M, Pitarch A, Sánchez M, Nombela C, Blackstock W, Gil C
Departamento de Microbiología II, Facultad de Farmacia, Universidad Complutense de Madrid, Spain.
Electrophoresis. 2000 Jul;21(13):2651-9. doi: 10.1002/1522-2683(20000701)21:13<2651::AID-ELPS2651>3.0.CO;2-3.
We have previously reported the usefulness of two-dimensional gel electrophoresis followed by Western blotting with sera from patients with systemic candidiasis in the detection of the major Candida albicans antigens (Pitarch et al., Electrophoresis 1999, 20, 1001-1010). The identification of these antigens would be useful for the characterization of good markers for the disease, and for the development of efficient diagnostic strategies. In this work we have used nanoelectrospray tandem mass spectrometry to obtain amino acid sequence information from the immunogenic proteins previously detected. We report here the cross-species identification of these antigens by matching of tandem mass spectrometry data to Saccharomyces cerevisiae proteins. Using this approach, we unambiguously identified the four C. albicans immunogenic proteins analyzed, namely aconitase, pyruvate kinase, phosphoglycerate mutase and methionine synthase. Furthermore, we report for the first time that aconitase, methionine synthase and phosphoglycerate mutase have antigenic properties in C. albicans.
我们之前曾报道过,二维凝胶电泳结合系统性念珠菌病患者血清进行蛋白质印迹分析,在检测白色念珠菌主要抗原方面具有实用性(皮塔尔奇等人,《电泳》,1999年,第20卷,第1001 - 1010页)。鉴定这些抗原对于确定该疾病的良好标志物以及开发有效的诊断策略将很有用处。在这项研究中,我们使用了纳米电喷雾串联质谱法来获取先前检测到的免疫原性蛋白质的氨基酸序列信息。我们在此报告通过将串联质谱数据与酿酒酵母蛋白质进行匹配,对这些抗原进行跨物种鉴定的情况。使用这种方法,我们明确鉴定出了所分析的四种白色念珠菌免疫原性蛋白质,即乌头酸酶、丙酮酸激酶、磷酸甘油酸变位酶和甲硫氨酸合酶。此外,我们首次报道乌头酸酶、甲硫氨酸合酶和磷酸甘油酸变位酶在白色念珠菌中具有抗原特性。