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Publication Details
AFRICAN RESEARCH NEXUS
SHINING A SPOTLIGHT ON AFRICAN RESEARCH
biochemistry, genetics and molecular biology
Kinetic properties of dihydrofolate reductase from wild-type and mutant Plasmodium vivax expressed in Escherichia coli
Molecular and Biochemical Parasitology, Volume 113, No. 2, Year 2001
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Description
Antifolate drugs inhibit malarial dihydrofolate reductase (DHFR). In Plasmodium falciparum, antifolate resistance has been associated with point mutations in the gene encoding DHFR. Recently, mutations at homologous positions have been observed in the P. vivax gene. Since P. vivax cannot be propagated in a continuous in vitro culture for drug sensitivity assays, the kinetic properties of DHFR were studied by expression of the DHFR domain in Escherichia coli. Induced expression yielded a protein product that precipitated as an inclusion body in E. coli. The soluble, active DHFR recovered after denaturation and renaturation was purified to homogeneity by affinity chromatography. Kinetic properties of the recombinant P. vivax DHFR showed that the wild-type DHFR (Ser-58 and Ser-117) and double mutant DHFR (Arg-58 and Asn-117) have similar Km values for dihydrofolate and NADPH. Antifolate drugs (pyrimethamine, cycloguanil, trimethoprim, and methotrexate), but not proguanil (parent compound of cycloguanil) inhibit DHFR activity, as expected. The kinetics of enzyme inhibition indicated that point mutations (Ser58Arg and Ser117Asn) are associated with lower affinity between the mutant enzyme and pyrimethamine and cycloguanil, which may be the origin of antifolate resistance. © 2001 Elsevier Science B.V.
Authors & Co-Authors
Tahar, Rachida
France, Paris
Cnrs Centre National de la Recherche Scientifique
de Pécoulas, Philippe Eldin
France, Paris
Cnrs Centre National de la Recherche Scientifique
Basco, Leonardo K.
France, Paris
Cnrs Centre National de la Recherche Scientifique
Cameroon, Yaounde
Inst. de Rech. Agric. Pour le Devmt.
Chiadmi, Mohammed
France, Paris
Cnrs Centre National de la Recherche Scientifique
Mazabraud, André R.
France, Paris
Cnrs Centre National de la Recherche Scientifique
France, Gif-sur-yvette
Université Paris-saclay
Statistics
Citations: 43
Authors: 5
Affiliations: 3
Identifiers
Doi:
10.1016/S0166-6851(01)00230-4
ISSN:
01666851
Research Areas
Genetics And Genomics