Aminopeptidase N1 (EtAPN1), an M1 Metalloprotease of the Apicomplexan Parasite Eimeria tenella, Participates in Parasite Development
Aminopeptidase N1 (EtAPN1), an M1 Metalloprotease of the Apicomplexan Parasite Eimeria tenella, Participates in Parasite Development
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DOI：
10.1128/ec.00062-14
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发表时间：
2014-07-01
期刊：
影响因子：
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通讯作者：
Brossier, Fabien
中科院分区：
文献类型：
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作者：
Gras, Simon;Byzia, Anna;Brossier, Fabien
氨基肽酶N是M1家族的金属蛋白酶，已在许多顶复体寄生虫中报道，包括疟原虫、弓形虫、隐孢子虫和艾美耳虫。在研究氨基肽酶作为球虫病（由艾美耳球虫引起的最重要的禽类疾病之一）治疗靶点的效力时，我们鉴定并鉴定了细艾美耳球虫氨基肽酶N1 （EtAPN1）。贝司他汀和阿马司他汀的抑制作用以及二价离子的再激活作用是典型的锌依赖性金属蛋白酶。EtAPN1与恶性疟原虫（PfA-M1）的a - m1具有相似的序列、三维结构、底物特异性和相似的动力学参数，后者是治疗疟疾的有效靶点。EtAPN1作为120 kda的前体合成，并在孢子形成过程中分裂成96、68和38 kda的形式。此外，免疫定位实验显示，与PfA-M1类似，EtAPN1在寄生虫细胞质和寄生虫细胞核的细胞内生命周期阶段都存在。目前的结果支持了两种氨基肽酶之间的保守作用的假设，我们认为EtAPN1可能是抗球虫药物的一个有价值的靶点。
Aminopeptidases N are metalloproteases of the M1 family that have been reported in numerous apicomplexan parasites, including Plasmodium, Toxoplasma, Cryptosporidium, and Eimeria. While investigating the potency of aminopeptidases as therapeutic targets against coccidiosis, one of the most important avian diseases caused by the genus Eimeria, we identified and characterized Eimeria tenella aminopeptidase N1 (EtAPN1). Its inhibition by bestatin and amastatin, as well as its reactivation by divalent ions, is typical of zinc-dependent metalloproteases. EtAPN1 shared a similar sequence, three-dimensional structure, and substrate specificity and similar kinetic parameters with A-M1 from Plasmodium falciparum (PfA-M1), a validated target in the treatment of malaria. EtAPN1 is synthesized as a 120-kDa precursor and cleaved into 96-, 68-, and 38-kDa forms during sporulation. Further, immunolocalization assays revealed that, similar to PfA-M1, EtAPN1 is present during the intracellular life cycle stages in both the parasite cytoplasm and the parasite nucleus. The present results support the hypothesis of a conserved role between the two aminopeptidases, and we suggest that EtAPN1 might be a valuable target for anticoccidiosis drugs.