Alvarado Magda E, Rubiano Claudia, Sánchez William, Díaz Andrea, Wasserman Moisés
Laboratorio de Investigaciones Básicas en Bioquímica - LIBBIQ, Facultad de Ciencias, Universidad Nacional de Colombia, Bogotá, Colombia.
Parasitol Res. 2018 Oct;117(10):3205-3214. doi: 10.1007/s00436-018-6019-z. Epub 2018 Jul 24.
Giardia intestinalis is a protozoan parasite that colonizes the upper part of the small intestine of its mammalian hosts. The trophozoite, which is the replicative stage, has a complex cytoskeleton that allows it to move and adhere to intestinal cells. It has been proposed that protein phosphatase 2A (PP2A) participates in the regulation of changes to the parasite cytoskeleton during its life cycle. However, how PP2A is involved in this regulation remains unclear since its substrates and regulators have not been characterized. In this work, we report the bioinformatic and experimental analysis of two potential regulatory B″ subunits of PP2A in Giardia, both of which are calcium-binding proteins. In this work, in silico and experimental evidence of the binding of both proteins to calcium is presented; the proteins are shown to interact with the catalytic PP2A subunit in the trophozoite stage, and they exhibit different subcellular localization patterns. Because PP2A is a heterotrimer, homology analysis of the different subunits of PP2A indicates that fewer holoenzyme combinations can be formed in this parasite than in other organisms. Our results suggest that the localization of PP2A may be associated with calcium-dependent signaling through its B″ type regulatory subunits.
肠贾第虫是一种原生动物寄生虫,寄生于哺乳动物宿主小肠上段。滋养体是其复制阶段,具有复杂的细胞骨架,使其能够移动并附着于肠道细胞。有人提出蛋白磷酸酶2A(PP2A)参与寄生虫生命周期中细胞骨架变化的调控。然而,由于其底物和调节因子尚未明确,PP2A如何参与这种调控仍不清楚。在这项研究中,我们报告了对贾第虫中PP2A两个潜在调节性B″亚基的生物信息学和实验分析,这两个亚基都是钙结合蛋白。在这项研究中,展示了这两种蛋白与钙结合的计算机模拟和实验证据;这些蛋白在滋养体阶段与催化性PP2A亚基相互作用,并且它们表现出不同的亚细胞定位模式。由于PP2A是一种异源三聚体,对PP2A不同亚基的同源性分析表明,与其他生物体相比,该寄生虫中可形成的全酶组合较少。我们的结果表明,PP2A的定位可能通过其B″型调节亚基与钙依赖性信号传导相关。