Confocal microscopic findings of cysteine protease calpain in Plasmodium falciparum |
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Authors: | Yun-Young Choi Suk-Yul Jung Pyo Yun Cho Bing Zheng Kie-In Park |
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Institution: | a Department of Infection Biology, Zoonosis Research Center, Wonkwang University School of Medicine, Chonbuk 570-749, South Korea b Department of Biomedical Laboratory Science, Namseoul University, 21 Maeju-ri, Seonghwan-eup, Choongnam 331-707, South Korea c College of Pharmacy, Wonkwang University, 344-2 Shinyong-dong, Iksan, Chonbuk 570-749, South Korea d Division of Biological Sciences, College of Natural Science, Chonbuk National University, Chonbuk 561-756, South Korea |
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Abstract: | Pf-calpain, a cysteine protease of Plasmodium falciparum, is believed to be one of the central mediators for essential parasitic activity. However, the roles of calpain on parasitic activity have not been determined in P. falciparum. In the present study, the localization of Pf-calpain was investigated using polyclonal antibodies (anti-Pf-calpain antibody A and B) against peptides that distinguished it from human calpain-7 and rat calpain-10 protein. Recombinant Pf-calpain (rPf-calpain) was identified as a 46 kDa protein using an anti-Pf-calpain antibody A, which can recognize the Pf-calpain binding site. Confocal microscopy revealed calpain within cytoplasmic localized parasites in the erythrocytic cycle. The findings suggested that the expression of Pf-calpain would be proportional to all different parasites in the erythrocytic cycle. On the other hand, anti-human calpain-7 antibody detected Pf-calpain in schizonts, and the immunofluorescence was stronger than with anti-rat calpain-10 antibody. However, the antibodies reacted with calpains in human red blood cells. These results show that anti-Pf-calpain antibody A and B specifically recognize only Pf-calpain. Taken together, the results suggest that Pf-calpain is expressed in all erythrocytic stages. In particular, the expression of Pf-calpain is increased much more when the late ring matures into the early trophozoite. Moreover, anti-Pf-calpain antibody A and B against synthetic peptides of the catalytic domain of Pf-calpain are useful to specifically detect Pf-calpain in all erythrocytic stages, while human and rat calpain antibody are not useful. |
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Keywords: | Pf-calpain Cysteine protease Plasmodium falciparum Confocal microscopy Erythrocytic cycle |
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