The MEP pathway in Babesia orientalis apicoplast, a potential target for anti-babesiosis drug development.
The MEP pathway in Babesia orientalis apicoplast, a potential target for anti-babesiosis drug development.
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东方巴贝虫顶端质体中的 MEP 途径，抗巴贝虫病药物开发的潜在靶标
DOI：
10.1186/s13071-018-3038-7
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发表时间：
2018-08-06
影响因子：
3.2
通讯作者：
Zhao J
中科院分区：
文献类型：
--
作者：
He L;He P;Luo X;Li M;Yu L;Guo J;Zhan X;Zhu G;Zhao J
中国水牛巴贝斯虫病的病原东方巴贝斯虫（Babesia orientalis）是一种顶复体寄生虫，在中国中南部地区广泛分布，每年造成巨大的经济损失。目前，还没有有效的疫苗或药物来对抗这种疾病。据报道，牛巴贝斯虫和恶性疟原虫的顶质体中含有磷米霉素可抑制的甲基赤藓糖醇磷酸（MEP）通路，提示该通路可作为筛选新药的靶点。然而，在东方桦中尚不为人所知。根据微小巴贝斯虫和牛巴贝斯虫的7个MEP通路基因设计引物。对这些基因进行克隆、测序和分析。将前两个酶基因1-脱氧-d -木糖5-磷酸合酶（BoDXS）和1-脱氧-d -木糖5-磷酸还原异构酶（BoDXR）的开放阅读框（ORFs）克隆到pET-32a表达载体上，表达为trx标签融合蛋白。制备兔抗rbodxs抗体和兔抗rbodxr抗体。采用Western blot方法鉴定了东方蓟中BoDXS和BoDXR的天然蛋白。采用Fosmidomycin和香叶香叶醇分别对东方白僵菌体外培养进行抑制试验和拯救试验。克隆并测序了东木MEP通路的7个酶基因（DXS、DXR、IspD、IspE、IspF、IspG和IspH），全长分别为2094、1554、1344、1521、654、1932和1056 bp。BoDXS和BoDXR表达为trx标签融合蛋白，大小分别为95和67 kDa。Western blot鉴定出原生BoDXS和原生BoDXR的条带分别为77 kDa和49 kDa。药物试验结果表明，fosmidomycin能抑制东方白僵菌的生长，香叶香叶醇能逆转fosmidomycin的作用。东方巴贝斯虫具有类异戊二烯生物合成途径，可能成为控制和治疗巴贝斯虫病的潜在药物靶点。鉴于fosmidomycin价格高且不稳定，进一步的研究应侧重于筛选稳定、廉价的药物。
The apicomplexan parasite Babesia orientalis, the causative agent of water buffalo babesiosis in China, is widespread in central and south China, resulting in a huge economic loss annually. Currently, there is no effective vaccine or drug against this disease. Babesia bovis and Plasmodium falciparum were reported to possess an apicoplast which contains the methylerythritol phosphate (MEP) pathway inhibitable by fosmidomycin, suggesting that the pathway could serve as a drug target for screening new drugs. However, it remains unknown in B. orientalis. Primers were designed according to the seven MEP pathway genes of Babesia microti and Babesia bovis. The genes were cloned, sequenced and analyzed. The open reading frames (ORFs) of the first two enzyme genes, 1-deoxy-D-xylulose 5-phosphate synthase (BoDXS) and 1-Deoxy-D-xylulose 5-phosphate reductoisomerase (BoDXR), were cloned into the pET-32a expression vector and expressed as a Trx-tag fusion protein. Rabbit anti-rBoDXS and rabbit anti-rBoDXR antibodies were generated. Western blot was performed to identify the native proteins of BoDXS and BoDXR in B. orientalis. Fosmidomycin and geranylgeraniol were used for inhibition assay and rescue assay, respectively, in the in vitro cultivation of B. orientalis. The seven enzyme genes of the B. orientalis MEP pathway (DXS, DXR, IspD, IspE, IspF, IspG and IspH) were cloned and sequenced, with a full length of 2094, 1554, 1344, 1521, 654, 1932 and 1056 bp, respectively. BoDXS and BoDXR were expressed as Trx-tag fusion proteins, with a size of 95 and 67 kDa, respectively. Western blot identified a 77 kDa band for the native BoDXS and a 49 kDa band for the native BoDXR. The drug assay results showed that fosmidomycin could inhibit the growth of B. orientalis, and geranylgeraniol could reverse the effect of fosmidomycin. Babesia orientalis has the isoprenoid biosynthesis pathway, which could be a potential drug target for controlling and curing babesiosis. Considering the high price and instability of fosmidomycin, further studies should focus on the screening of stable and cheap drugs.
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影响因子：
5.2
作者：
Imlay, Leah;Odom, Audrey R
通讯作者：
Odom, Audrey R
影响因子：
1.5
作者：
Odom, Audrey R.;Van Voorhis, Wesley C.
通讯作者：
Van Voorhis, Wesley C.
DOI：
10.1093/bioinformatics/btw108
发表时间：
2016-07-01
期刊：
Bioinformatics (Oxford, England)
影响因子：
--
作者：
Katoh K;Standley DM
通讯作者：
Standley DM
影响因子：
1.5
作者：
Caballero, Marina C.;Pedroni, Monica J.;Palmer, Guy H.;Suarez, Carlos E.;Davitt, Christine;Lau, Audrey O. T.
通讯作者：
Lau, Audrey O. T.
影响因子：
4.9
作者：
Lizundia, Regina;Werling, Dirk;Ralph, Stuart A.
通讯作者：
Ralph, Stuart A.