Laboratório de Interação Parasito-Hospedeiro, Faculdade de Medicina, Universidade de Brasília, 70910-900-DF, Brazil.
Microbes Infect. 2010 Jun;12(6):457-66. doi: 10.1016/j.micinf.2010.02.007. Epub 2010 Feb 24.
Proteases play important roles in many biological processes of parasites, including their host interactions. In sleeping sickness, Trypanosoma brucei proteases released into the host bloodstream could hydrolyze host factors, such as hormones, contributing to the development of the disease's symptoms. In this study, we present the identification of the T. brucei prolyl oligopeptidase gene (poptb) and the characterization of its corresponding enzyme, POP Tb. Secondary structure predictions of POP Tb show a structural composition highly similar to other POPs. Recombinant POP Tb produced in E. coli was active and highly sensitive to inhibitors of Trypanosoma cruzi POP Tc80. These inhibitors, which prevent T. cruzi entry into non-phagocytic cells, arrested growth of the T. brucei bloodstream form in a dose-dependent manner. POP Tb hydrolyzes peptide hormones containing Pro or Ala at the P1 position at a slightly alkaline pH, and also cleaves type I collagen in vitro and native collagen present in rat mesentery. Furthermore, POP Tb is released into the bloodstream of T. brucei infected mice where it remains active. These data suggest that POP Tb might contribute to the pathogenesis of sleeping sickness.
蛋白酶在寄生虫的许多生物学过程中发挥着重要作用,包括它们与宿主的相互作用。在昏睡病中,释放到宿主血液中的布氏锥虫蛋白酶可以水解宿主因子,如激素,导致疾病症状的发展。在这项研究中,我们鉴定了布氏锥虫脯氨酰寡肽酶基因(poptb)及其相应酶 POP Tb 的特性。POP Tb 的二级结构预测显示其结构组成与其他 POPs 高度相似。在大肠杆菌中产生的重组 POP Tb 具有活性,并对抑制克氏锥虫 POP Tc80 的抑制剂高度敏感。这些抑制剂可阻止克氏锥虫进入非吞噬细胞,以剂量依赖的方式阻止布氏锥虫血液形式的生长。POP Tb 在略微碱性的 pH 下水解 P1 位含有 Pro 或 Ala 的肽激素,并且还在体外切割 I 型胶原蛋白和大鼠肠系膜中存在的天然胶原蛋白。此外,POP Tb 被释放到感染布氏锥虫的小鼠的血液中,在那里它仍然保持活性。这些数据表明,POP Tb 可能有助于昏睡病的发病机制。