Reyes-Uribe Patrícia, Pereira-dos-Santos Taissa, De Jesus Jose B, Mesquita-Rodrigues Camila, Arevalo Jorge, Cupolillo Elisa, Cuervo Patricia
Laboratório de Biologia Molecular de Tripanosomatídeos, Universidad Peruana Cayetano Heredia, Lima, Peru.
Parasitol Int. 2012 Dec;61(4):513-9. doi: 10.1016/j.parint.2012.03.007. Epub 2012 Mar 30.
American tegumentary leishmaniasis (ATL) in Peru is mainly associated with Leishmania (Viannia) peruviana and L. (V.) braziliensis. These parasites are genetically related, and their characterization as distinct species is controversial. Despite their genetic similarity, each species is associated with different clinical manifestations of ATL; L. (V.) peruviana causes only cutaneous leishmaniasis, whereas L. (V.) braziliensis can cause both cutaneous and mucocutaneous leishmaniasis. Because the primary cutaneous lesions caused by infection with these species are indistinguishable, it is necessary to develop a suitable method to differentiate them in order to prevent possible metastasis to oropharyngeal mucosa. In the present study, we investigated the proteolytic profile of L. (V.) peruviana and L. (V.) braziliensis isolates from Peru by zymographic analysis in SDS-PAGE copolymerized with gelatin. Enzymes were characterized according to their pH range of activity and sensitivity to distinct peptidase inhibitors. We observed that L. (V.) peruviana isolates displayed three proteolytic bands with molecular masses ranging from 55 to 80 kDa, whereas L. (V.) braziliensis isolates showed six proteolytic activities between 55 and 130 kDa. Using specific inhibitors, we determined that these proteolytic activities are due to metallopeptidases and present optimal activity between the pH range 5.5 and 10.0. Our results suggest that the expression of metallopeptidases in L. (V.) peruviana and L. (V.) braziliensis isolates from Peru is species-specific.
秘鲁的美洲皮肤利什曼病（ATL）主要与秘鲁利什曼原虫（维氏亚属）和巴西利什曼原虫（维氏亚属）有关。这些寄生虫在基因上相关，将它们鉴定为不同物种存在争议。尽管它们在基因上相似，但每个物种都与ATL的不同临床表现相关；秘鲁利什曼原虫（维氏亚属）仅引起皮肤利什曼病，而巴西利什曼原虫（维氏亚属）可引起皮肤和黏膜皮肤利什曼病。由于这些物种感染引起的原发性皮肤病变无法区分，因此有必要开发一种合适的方法来区分它们，以防止可能转移至口咽黏膜。在本研究中，我们通过在与明胶共聚的SDS-PAGE中进行酶谱分析，研究了来自秘鲁的秘鲁利什曼原虫（维氏亚属）和巴西利什曼原虫（维氏亚属）分离株的蛋白水解谱。根据酶的活性pH范围和对不同肽酶抑制剂的敏感性对酶进行了表征。我们观察到，秘鲁利什曼原虫（维氏亚属）分离株显示出三条分子量在55至80 kDa之间的蛋白水解带，而巴西利什曼原虫（维氏亚属）分离株在55至130 kDa之间显示出六种蛋白水解活性。使用特异性抑制剂，我们确定这些蛋白水解活性是由金属肽酶引起的，并且在pH范围5.5至10.0之间呈现最佳活性。我们的结果表明，来自秘鲁的秘鲁利什曼原虫（维氏亚属）和巴西利什曼原虫（维氏亚属）分离株中金属肽酶的表达具有物种特异性。
