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Elongation factor 1-alpha is released into the culture medium during growth of Giardia intestinalis trophozoites

机译:贾第鞭毛虫滋养体生长过程中将伸长因子1-α释放到培养基中

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The molecular pathogenesis of the intestinal parasite Giardia intestinalis is still not fully understood but excretory-secretory products have been suggested to be important during host-parasite interactions. Here we used SOS-PAGE gels and MALDI-TOF analysis to identify proteins released by Giardia trophozoites during in vitro growth. Serum proteins (mainly bovine serum albumin) in the growth medium, bind to the parasite surface and they are continuously released, which interfere with parasite secretome characterization. However, we identified two released Giardia proteins: elongation factor-1 alpha (EF-1 alpha) and a 58 kDa protein, identified as arginine deiminase (ADI). This is the first description of EF-1 alpha as a released/secreted Giardia protein, whereas ADI has been identified in an earlier secretome study. Two genes encoding EF-1 alpha were detected in the Giardia WB genome 35 kbp apart with almost identical coding sequences but with different promoter and 3' regions. Promoter luciferase-fusions showed that both genes are transcribed in trophozoites. The EF-1 alpha protein localizes to the nuclear region in trophozoites but it relocalizes to the cytoplasm during host-cell interaction. Recombinant EF-1 alpha is recognized by serum from giardiasis patients. Our results suggest that released EF-1 alpha protein can be important during Giardia infections
机译:肠道寄生虫贾第鞭毛虫的分子发病机制仍未完全了解,但已建议在宿主-寄生虫相互作用期间排泄-分泌产物很重要。在这里,我们使用SOS-PAGE凝胶和MALDI-TOF分析来鉴定贾第鞭毛虫滋养体在体外生长过程中释放的蛋白质。生长培养基中的血清蛋白(主要是牛血清白蛋白)与寄生虫表面结合并不断释放,这会干扰寄生虫分泌组的鉴定。但是,我们鉴定了两种释放的贾第虫蛋白:延伸因子1α(EF-1 alpha)和58 kDa蛋白,被鉴定为精氨酸脱亚氨酶(ADI)。这是EF-1α作为释放/分泌的贾第鞭毛虫蛋白的首次描述,而ADI在早期的分泌组研究中已被鉴定。在相距35 kbp的贾第鞭毛虫WB基因组中检测到两个编码EF-1 alpha的基因,它们的编码序列几乎相同,但启动子和3'区不同。启动子荧光素酶融合显示这两个基因都在滋养体中转录。 EF-1α蛋白位于滋养体中的核区,但在宿主细胞相互作用期间重新定位到细胞质。贾第鞭毛虫病患者的血清可识别重组EF-1α。我们的结果表明,释放的EF-1α蛋白在贾第虫感染期间可能很重要

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