H2O2 dynamics in the malaria parasite Plasmodium falciparum
H2O2 dynamics in the malaria parasite Plasmodium falciparum
复制标题
DOI：
10.1371/journal.pone.0174837
复制
发表时间：
2017-04-03
期刊：
影响因子：
3.7
通讯作者：
Becker, Katja
中科院分区：
文献类型：
--
作者：
Rahbari, Mahsa;Rahlfs, Stefan;Becker, Katja
过氧化氢是一种重要的抗菌剂，但也是至关重要的参与氧化还原信号和病原体-宿主细胞相互作用。作为系统研究活疟原虫细胞内H2 O2动力学和调节的基础，我们在恶性疟原虫中建立了基因编码的荧光H2 O2传感器roGFP 2-Orp 1和HyPer-3。这两个比例氧化还原探针以及pH控制SypHer在血阶段寄生虫的胞质溶胶中表达。这两种氧化还原传感器显示出可重复的敏感性，对H2 O2在较低的微摩尔范围内，在体外和寄生虫。由于HyPer-3的pH敏感性，我们使用表达roGFP 2-Orp 1的寄生虫来评估抗疟药物对疟原虫中H2 O2水平和解毒的短期、中期和长期影响。没有喹啉类或青蒿素测试有可检测到的直接影响H2 O2的稳态，在相关的浓度。然而，用抗疟药物或热休克预处理细胞导致对外源性H2 O2的更高耐受性。系统的评价和比较的两个基因编码的胞质H2 O2探针在疟疾寄生虫提供了一个基础，研究寄生虫-宿主细胞相互作用或药物的影响与时空分辨率，同时保持细胞的完整性。
Hydrogen peroxide is an important antimicrobial agent but is also crucially involved in redox signaling and pathogen-host cell interactions. As a basis for systematically investigating intracellular H2O2 dynamics and regulation in living malaria parasites, we established the genetically encoded fluorescent H2O2 sensors roGFP2-Orp1 and HyPer-3 in Plasmodium falciparum. Both ratiometric redox probes as well as the pH control SypHer were expressed in the cytosol of blood-stage parasites. Both redox sensors showed reproducible sensitivity towards H2O2 in the lower micromolar range in vitro and in the parasites. Due to the pH sensitivity of HyPer-3, we used parasites expressing roGFP2-Orp1 for evaluation of short-, medium-, and long-term effects of antimalarial drugs on H2O2 levels and detoxification in Plasmodium. None of the quinolines or artemisinins tested had detectable direct effects on the H2O2 homeostasis at pharmacologically relevant concentrations. However, pre-treat-ment of the cells with antimalarial drugs or heat shock led to a higher tolerance towards exogenous H2O2. The systematic evaluation and comparison of the two genetically encoded cytosolic H2O2 probes in malaria parasites provides a basis for studying parasite-host cell interactions or drug effects with spatio-temporal resolution while preserving cell integrity.