Glycerol biosynthetic pathway plays an essential role in proliferation and antioxidative defense in the human enteric protozoan parasite Entamoeba histolytica.
Glycerol biosynthetic pathway plays an essential role in proliferation and antioxidative defense in the human enteric protozoan parasite Entamoeba histolytica.
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DOI：
10.1038/s41598-023-40670-z
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发表时间：
2023-09-05
影响因子：
4.6
通讯作者：
中科院分区：
文献类型：
--
作者：
阿米巴病由原生动物寄生虫溶组织内阿米巴引起。除甲硝唑及其衍生物外的治疗选择很少，它们对无症状囊肿携带者的疗效较低，体外耐药性的实验证据证明了发现/重新利用抗阿米巴病新药的运动。大肠杆菌对氧化应激和半胱氨酸剥夺的整体代谢反应。histolytica揭示甘油代谢可能代表药物开发的合理靶点。在这项研究中，使用14 C标记的葡萄糖，只有11%的总葡萄糖被E。溶组织滋养体与脂质结合。为了更好地了解甘油代谢在这种寄生虫中的作用，我们专注于表征两种重要的酶，甘油激酶（GK）和甘油3-磷酸脱氢酶（G3 PDH）。重组GK进行了详细的生化特性，而G3 PDH不是由于蛋白表达和纯化的失败。GK在逆反应中表现出了前所未有的动力学特性。基因沉默表明，GK是必不可少的最佳生长，而G3 PDH不是。RNA-seq分析显示，G3 PDH基因沉默导致GK表达上调，而GK基因沉默导致抗氧化酶表达上调。虽然GK和抗氧化酶的上调之间的精确的分子联系没有得到证实，GK基因沉默后抗氧化酶表达的增加表明GK和细胞对氧化应激的反应之间存在潜在的联系。总之，这些结果提供了第一个直接证据的生物重要性和协调调节甘油代谢途径的增殖和抗氧化防御在大肠杆菌。histolytica，证明开发这些酶作为未来的药物靶点。
Amebiasis is caused by the protozoan parasite Entamoeba histolytica. Treatment options other than metronidazole and its derivatives are few, and their low efficacy against asymptomatic cyst carriers, and experimental evidence of resistance in vitro justify the discovery/repurposing campaign for new drugs against amebiasis. Global metabolic responses to oxidative stress and cysteine deprivation by E. histolytica revealed glycerol metabolism may represent a rational target for drug development. In this study using 14C-labelled glucose, only 11% of the total glucose taken up by E. histolytica trophozoites is incorporated to lipids. To better understand the role of glycerol metabolism in this parasite, we focused on characterizing two important enzymes, glycerol kinase (GK) and glycerol 3-phosphate dehydrogenase (G3PDH). Recombinant GK was biochemically characterized in detail, while G3PDH was not due to failure of protein expression and purification. GK revealed novel characteristics and unprecedented kinetic properties in reverse reaction. Gene silencing revealed that GK is essential for optimum growth, whereas G3PDH is not. Gene silencing of G3PDH caused upregulated GK expression, while that of GK resulted in upregulation of antioxidant enzymes as shown by RNA-seq analysis. Although the precise molecular link between GK and the upregulation of antioxidant enzymes was not demonstrated, the observed increase in antioxidant enzyme expression upon GK gene silencing suggests a potential connection between GK and the cellular response to oxidative stress. Together, these results provide the first direct evidence of the biological importance and coordinated regulation of the glycerol metabolic pathways for proliferation and antioxidative defense in E. histolytica, justifying the exploitation of these enzymes as future drug targets.
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影响因子：
3.7
作者：
Jeelani G;Sato D;Husain A;Escueta-de Cadiz A;Sugimoto M;Soga T;Suematsu M;Nozaki T
通讯作者：
Nozaki T
影响因子：
6.7
作者：
Bracha R;Nuchamowitz Y;Anbar M;Mirelman D
通讯作者：
Mirelman D
影响因子：
14.8
作者：
Kelley LA;Mezulis S;Yates CM;Wass MN;Sternberg MJ
通讯作者：
Sternberg MJ
DOI：
10.1016/j.ijpddr.2018.02.004
发表时间：
2018-04
期刊：
International journal for parasitology. Drugs and drug resistance
影响因子：
--
作者：
Nurkanto A;Jeelani G;Yamamoto T;Naito Y;Hishiki T;Mori M;Suematsu M;Shiomi K;Hashimoto T;Nozaki T
通讯作者：
Nozaki T
影响因子：
12.3
作者：
Kircher M;Stenzel U;Kelso J
通讯作者：
Kelso J