Drug uptake and modulation of drug resistance in Leishmania by an aquaglyceroporin
Drug uptake and modulation of drug resistance in Leishmania by an aquaglyceroporin
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DOI：
10.1074/jbc.m403959200
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发表时间：
2004-07-23
影响因子：
4.8
通讯作者：
Mukhopadhyay, R
中科院分区：
文献类型：
--
作者：
Gourbal, B;Sonuc, N;Mukhopadhyay, R
利什曼病是一种原生动物寄生虫病，影响全世界1200万人。治疗本病的首选药物是锑剂。在流行地区，对这类药物的抗药性是治疗的主要障碍。微生物通常通过摄取系统的突变或下调而对药物产生耐药性，但利什曼原虫对锑药物的摄取系统尚不清楚。在其他生物体中，水甘油孔蛋白已被证明有助于三价类金属的吸收。在这项研究中，我们报告的鉴定和表征的水甘油孔蛋白从利什曼原虫主要（LmAQP1）和利什曼原虫tarentolae（LtAQP1），分别。这些利什曼原虫蛋白具有水甘油孔蛋白的保守特征基序。将LmAQP1基因转染到三种利什曼原虫中，L. tarentolae、婴儿利什曼原虫（Leishmania infantum）和L.主要的，产生过敏的As（III）和Sb（III）在所有三个菌株。免疫印迹法检测到LmAQP1的产生增加。具有导致耐药机制的各种突变的耐药寄生虫在LmAQP1表达后对两种金属过敏。As（III）或Sb（III）的摄取率增加与野生型和耐药转染子的类金属敏感性相关。在Pentostam耐药的田间分离株中转染LmAQP1也致敏了巨噬细胞相关无鞭毛体形式的寄生虫。LmAQP1的一个等位基因在L. major，由此产生的细胞对Sb（III）的抗性提高了10倍。这是利什曼原虫中aquaglyceroporin吸收类金属药物的第一份报告，表明了在该领域逆转耐药性的策略。
Leishmaniasis is a protozoan parasitic disease that affects 12 million people worldwide. The first line choice for the treatment of this disease is antimonial drugs. In the endemic regions, resistance to this class of drugs is a major impediment to treatment. Microbes often become resistant to drugs by mutation or down-regulation of uptake systems, but the uptake system for the antimonial drugs in Leishmania is unknown. In other organisms, aquaglyceroporins have been shown to facilitate uptake of trivalent metalloids. In this study, we report the identification and characterization of aquaglyceroporins from Leishmania major (LmAQP1) and Leishmania tarentolae (LtAQP1), respectively. These Leishmania proteins have the conserved signature motifs of aquaglyceroporins. Transfection of LmAQP1 into three species of Leishmania, L. tarentolae, Leishmania infantum, and L. major, produced hypersensitivity to both As(III) and Sb(III) in all three strains. Increased production of LmAQP1 was detected by immunoblotting. Drug-resistant parasites with various mutations leading to resistance mechanisms became hypersensitive to both metalloids after expression of LmAQP1. Increased rates of uptake of As( III) or Sb( III) correlated with metalloid sensitivity of the wild type and drug-resistant transfectants. Transfection of LmAQP1 in a Pentostam-resistant field isolate also sensitized the parasite in the macrophage-associated amastigote form. One allele of LmAQP1 was disrupted in L. major, and the resulting cells became 10-fold more resistant to Sb( III). This is the first report of the uptake of a metalloid drug by an aquaglyceroporin in Leishmania, suggesting a strategy to reverse resistance in the field.