抗日本血吸虫生殖抗原SIEA 26-28 ku组分的质谱分析

目的 筛选确定抗日本血吸虫生殖功能抗原SIEA26-28 ku的编码基因. 方法 采用固相pH梯度双向凝胶电泳分离日本血吸虫未成熟卵可溶性抗原(SIEA),选择SIEA 26-28 ku免疫血清识别信号较强的SIEA-2D蛋白质斑点(65～73号)采样,消化处理后进行质谱与生物学信息分析. 结果 从SIEA-2D图谱上共获得1 037个蛋白质斑点;通过MALDI-TOF-MS对其进行肽指纹图谱分析后获得了7张肽指纹图谱.通过PeptIdent软件检索SWISS-PROT数据库,发现有意义的血吸虫同源蛋白质11个,这些同源蛋白质部分与细胞代谢或蛋白质合成代谢相关,部分与核苷酸代谢有关,其中编号为SIEA-2D66为日本血吸虫次黄嘌呤-鸟嘌呤磷酸核糖转移酶(HGPRT),SIEA-2D71为琥珀酸脱氢酶铁硫蛋白(SDISP),SIEA-2D73为谷胱甘肽-S-转移酶(GST),其同源程度分别为37.2%、18.7%和22.9%. 结论 初步获得与抗日本血吸虫生殖功能抗原SIEA 26～28 ku相关的编码基因HGPRT、SDISP和GST.

MALDI-TOF-MS analysis of the SIEA 26-28 ku components against Schisitosoma japonicum fecundity immunity

Objective To identify and get the encoding gene of the SIEA 26-28 ku molecule against Schisitosoma japonicum fecundity immunity. Methods The proteome group of SIEA was separated and identified by a series of methods, including immobilized pH gradient two-dimensional polyacrylamide gel electrophoresis (2-D), silver staining etc. Spots of SIEA-2D65-73, which showed a strong immunoreactivity with the mono-specific anti-sera against SIEA26-28 ku components, were selected and incised from silver staining gel after digested in-gel by trypsin, and used to analyzed by the matrix-assisted laser desorption/ionization time of flying mass spectrometry (MALDI-TOF-MS) and bioinformatics analysis.Results The silver stained 2-DE profile of SIEA demonstrated 1 037 protein spots. Seven peptide mass fingerprint (PMF) maps were displayed by MALDI-TOF-MS. The typical peptide masses were searched from the SWISS-PROT database with PeptIdent software. Eleven proteins with significant homogeneity were preliminarily identified, which included cell metabolism-related proteins, enzymes related to the proteins syntheses, and another of them were responsible for the salvage pathway of purine bases needed for nucleotide metabolism. Three of these were preliminarily identified proteins including the SIEA-2D66 (hypoxanthine-guanine phosphoribosyl transferase, HGPRT), SIEA-2D71 (succinate dehydrogenase iron-sulfur protein, SDISP) and SIEA-2D73 (glutathione-S-transferase, GST) shared a homogeneity of 37.2%, 18.7% and 22.9%, respectively. Conclusion HGPRT, SDISP and GST are relative to the encoding gene of SIEA 26-28 ku molecule against S. japonicum fecundity immunity.