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Characterization of PFF1010c, a type IV Plasmodium fasciparum heat shock protein 40Mutavhatsindi, Hygon January 2016 (has links)
MSc (Biochemistry) / Department of Biochemistry / Malaria is caused by protozoa of the genus Plasmodium. Malaria accounts for approximately more
than half a million deaths yearly. Of the five species of Plasmodium, P. falciparum accounts for
the most deadly form of the disease. P. falciparum survives under various physiological conditions
during its life cycle. The parasite employs its molecular chaperones machinery particularly heat
shock proteins (Hsps) to protect its protein constituents during physiological stress. Hsps are
conserved molecules that constitute a major part of the cell’s molecular chaperone system. P.
falciparum Hsps play an important cyto-protective role guaranteeing that the malarial parasite
survives under the severe conditions that prevail in the host environment. PFF1010c is a type
IV P. falciparum heat shock protein 40. PFF1010c is predicted to be expressed only at the
gametocyte stage of the malarial parasite’s life cycle. The aim of the current study was to
investigate the expression PFF1010c by parasites and the gametocyte stage as well as characterize
the structure-function features of the protein. PFF1010c was successfully expressed in E. coli cells.
Despite successful expression of the protein, its purification proved problematic. The attempt to
purify PFF1010c was carried out under both native and denaturing conditions. Far Western blot
analysis to investigate direct interaction between PFF1010c and PfHsp70-1 was conducted and no
interaction was observed. Malarial parasites were harvested at different stages and total protein
was isolated. The expression of PFF1010c was confirmed to occur at the gametocyte stage of the
parasite’s development using Western blot analysis. This study confirmed that PFF1010c is only
expressed at the gametocyte stage of the malarial parasite. Furthermore, PFF1010c was not
expressed at the asexual stage. Possible interactors of PFF1010c were predicted by STRING, a
bioinformatics based tool. The expression of PfHsp90, PfHop and PfHsp70-1 at the gametocyte
stage was investigated and confirmed by Western blot analyses.
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