Functional characterization of an alternative [lactate dehydrogenase-like] malate dehydrogenase in Plasmodium falciparum
- 1 January 2004
- journal article
- Published by Springer Science and Business Media LLC in Zeitschrift für Parasitenkunde
- Vol. 92 (1), 43-47
- https://doi.org/10.1007/s00436-003-0996-1
Abstract
The catalysis of malate dehydrogenase (MDH) in Plasmodium falciparum (pfMDH) which involves NAD/NADH coupling is crucial for the parasite’s pathogenicity. Primers were designed based on the P. falciparum genome resource, and these facilitated the cloning of a gene coding for pfMDH from a local clinical isolate. The DNA sequence of the cloned gene revealed an open-reading frame that encodes a protein of 313 amino acids. After induction in Escherichia coli BL21, enzyme assays of the expressed pfMDH purified by affinity chromatography exhibited significant enzyme activity of about 50 U/mg, where one unit (U) of enzyme activity is defined as the amount of enzyme oxidising 1 μol NADH/min. Based on its phylogenetic status amongst MDHs and lactate dehydrogenases (LDHs), the cloned gene was clearly defined as belonging to the NADH-dependent [LDH-like] MDHs. It is noteworthy that pfMDH harbours unique structural characteristics potentially useful for screening drugs specific for disabling parasitic enzymes.Keywords
This publication has 10 references indexed in Scilit:
- Genome sequence of the human malaria parasite Plasmodium falciparumNature, 2002
- A proteomic view of the Plasmodium falciparum life cycleNature, 2002
- Molecular Evolution Within the L-Malate and L-Lactate Dehydrogenase Super-FamilyJournal of Molecular Evolution, 2002
- Structural basis of substrate specificity in malate dehydrogenases: crystal structure of a ternary complex of porcine cytoplasmic malate dehydrogenase, α-Ketomalonate and TetrahydoNADJournal of Molecular Biology, 1999
- Malate dehydrogenase: A model for structure, evolution, and catalysisProtein Science, 1994
- Expression of Plasmodium falciparum lactate dehydrogenase in Escherichia coliMolecular and Biochemical Parasitology, 1993
- Malate dehydrogenase isoenzymes: Cellular locations and role in the flow of metabolites between the cytoplasm and cell organellesBiochimica et Biophysica Acta (BBA) - Bioenergetics, 1992
- Purification and properties of Plasmodium falciparum malate dehydrogenaseMolecular and Biochemical Parasitology, 1992
- Glycolytic pathway of the human malaria parasite Plasmodium falciparum: primary sequence analysis of the gene encoding 3-phosphoglycerate kinase and chromosomal mapping studiesGene, 1991
- Cloning metabolic pathway genes by complementation in Escherichia coli. Isolation and expression of Plasmodium falciparum glucose phosphate isomerase.Journal of Biological Chemistry, 1990