Departamento de Biología Molecular y Biotecnología, Universidad Nacional Autónoma de México, Apartado Postal 70-228, México, DF, Mexico.
Exp Parasitol. 2011 Jan;127(1):249-59. doi: 10.1016/j.exppara.2010.08.003. Epub 2010 Aug 9.
The expression and biological role of actin during the Trypanosoma cruzi life cycle remains largely unknown. Polyclonal antibodies against a recombinant T. cruzi actin protein were used to confirm its expression in epimastigotes, trypomastigotes, and amastigotes. Although the overall levels of expression were similar, clear differences in the subcellular distribution of actin among the developmental stages were identified. The existence of five actin variants in each developmental stage with distinct patterns of expression were uncovered by immunoblotting of protein extracts separated 2D-SDS gels. The isoelectric points of the actin variants in epimastigotes ranged from 4.45 to 4.9, whereas they ranged from 4.9 to 5.24 in trypomastigotes and amastigotes. To determine if the actin variants found could represent previously unidentified actins, we performed a genomic survey of the T.cruzi GeneDB database and found 12 independent loci encoding for a diverse group of actins and actin-like proteins that are conserved among trypanosomatids.
在克氏锥虫生命周期中,肌动蛋白的表达和生物学作用在很大程度上仍是未知的。使用针对重组克氏锥虫肌动蛋白蛋白的多克隆抗体来确认其在滋养体、动基体和无鞭毛体中的表达。尽管表达的总体水平相似,但在不同发育阶段中肌动蛋白的亚细胞分布存在明显差异。通过对分离的 2D-SDS 凝胶中的蛋白提取物进行免疫印迹,发现了每个发育阶段中存在五种具有不同表达模式的肌动蛋白变体。滋养体中肌动蛋白变体的等电点范围为 4.45 至 4.9,而在动基体和无鞭毛体中范围为 4.9 至 5.24。为了确定所发现的肌动蛋白变体是否可以代表以前未鉴定的肌动蛋白,我们对 T.cruzi GeneDB 数据库进行了基因组调查,发现了 12 个独立的基因座,编码了一组多样化的肌动蛋白和肌动蛋白样蛋白,这些蛋白在原生动物中是保守的。