tailieunhanh - Báo cáo khoa học: Isocitrate dehydrogenase of Plasmodium falciparum Energy metabolism or redox control?

Erythrocytic stages of the malaria parasitePlasmodium fal-ciparumrely on glycolysis for their energy supply and it is unclear whether they obtain energy via mitochondrial res-piration albeit enzymes of the tricarboxylic acid (TCA) cycle appear to be expressed in these parasite stages. Isocitrate dehydrogenase (ICDH) is either an integral part of the mitochondrial TCA cycle or is involved in providing NADPH for reductive reactions in the cell. | Eur. J. Biochem. 270 1775-1783 2003 FEBS 2003 doi Isocitrate dehydrogenase of Plasmodium falciparum Energy metabolism or redox control Carsten Wrenger and Sylke Miiller Division of Biological Chemistry and Molecular Microbiology School of Life Sciences University of Dundee UK Erythrocytic stages of the malaria parasite Plasmodium falciparum rely on glycolysis for their energy supply and it is unclear whether they obtain energy via mitochondrial respiration albeit enzymes of the tricarboxylic acid TCA cycle appear to be expressed in these parasite stages. Isocitrate dehydrogenase ICDH is either an integral part of the mitochondrial TCA cycle or is involved in providing NADPH for reductive reactions in the cell. The gene encoding P. facciparum ICDH was cloned and analysis of the deduced amino-acid sequence revealed that it possesses a putative mitochondrial targeting sequence. The protein is very similar to NADP -dependent mitochondrial counterparts of higher eukaryotes but not Escherichia coli. Expression of full-length ICDH generated recombinant protein exclusively expressed in inclusion bodies but the removal of 27 N-terminal amino acids yielded appreciable amounts of soluble ICDH consistent with the prediction that these residues confer targeting of the native protein to the parasites mitochondrion. Recombinant ICDH forms homodimers of 90 kDa and its activity is dependent on the bivalent metal ions Mg2 or Mn2 with apparent Km values of 13 M and 22 M respectively. Plasmodium ICDH requires NADP as cofactor and no activity with NAD was detectable the Kmpp for NADP was found to be 90 M and that of D-isocitrate was determined to be 40 M. Incubation of under exogenous oxidative stress resulted in an up-regulation of ICDH mRNA and protein levels indicating that the enzyme is involved in mitochondrial redox control rather than energy metabolism of the parasites. Keywords isocitrate dehydrogenase redox control mitochondrion .

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