Falcipain-1, a Plasmodium falciparum cysteine protease with vaccine potential

Chauhan, Virander Singh ; Kumar, Amit ; Kumar, Krishan ; Korde, Reshma ; Puri, Sunil Kumar ; Malhotra, Pawan (2007) Falcipain-1, a Plasmodium falciparum cysteine protease with vaccine potential Infection and Immunity, 75 (4). pp. 2026-2034. ISSN 0019-9567

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Official URL: http://iai.asm.org/cgi/content/abstract/75/4/2026

Related URL: http://dx.doi.org/10.1128/IAI.01533-06

Abstract

Cysteine proteases (falcipains) of Plasmodium falciparum are potential targets for antimalarial chemotherapy, since they have been shown to be involved in important cellular functions such as hemoglobin degradation and invasion/rupture of red blood cells during parasite life cycle. The role of falcipain-1 at the asexual blood stages of the parasite still remains uncertain. This is mainly due to a lack of methods to prepare this protein in an active form. In order to obtain biologically active falcipain-1, a number of falcipain-1 constructs were designed and a systematic assessment of the refolding conditions was done. We describe here the expression, purification, and characterization of a falcipain-1 construct encoding mature falcipain-1 and 35 amino acids from the C-terminal of the pro domain. Recombinant falcipain-1 was overexpressed in the form of inclusion bodies, solubilized, and purified by Ni2+-nitrilotriacetic acid affinity chromatography under denaturing conditions. A systemic approach was then followed to optimize refolding parameters. An optimum refolding condition was obtained, and the yield of the purified refolded falcipain-1 was ~1 mg/liter. Activity of the protein was analyzed by fluorometric and gelatin degradation assays. Immunolocalization studies using anti-falcipain-1 sera revealed a distinct staining at the apical end of the P. falciparum merozoites. Previous studies using falcipain-1-specific inhibitors have suggested a role of falcipain-1 in merozoite invasion. Based on its localization and its role in invasion, we analyzed the immunogenicity of falcipain-1 in mice, followed by heterologous challenge with Plasmodium yoelii sporozoites. Our results suggest a possible role of falcipain-1 in merozoite invasion.

Item Type:Article
Source:Copyright of this article belongs to American Society for Microbiology.
ID Code:8278
Deposited On:26 Oct 2010 11:52
Last Modified:16 May 2016 18:18

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