Molecular analysis of Plasmodium falciparum hexokinase.
Hexokinase, a key glycolytic enzyme, is involved in the initial phosphorylation reaction of imported glucose and specific blocking of this activity may therefore arrest the development of malaria parasites. We describe here the cloning of a single copy hexokinase gene of Plasmodium falciparum (PfHK) from cDNA or genomic DNA libraries. The deduced amino acid sequence of PfHK has 26% identity with human hexokinase I and its predicted molecular mass assigns it as an invertebrate type isoenzyme of hexokinase. A single 1.5-kb exon is translated from a 3-kb mRNA in asexual stages of the parasite. In contrast to aldolase and GPI, the gene for this glycolytic enzyme is located on chromosome 8. Poly- and monoclonal antibodies against recombinant PfHK support our cloning results at the protein level as they detect a protein of the predicted size and isoelectric point by Western blotting in parasite protein samples. Moreover, polyclonal rabbit IgG against recombinant PfHK partially inhibits the hexokinase activity of a P. falciparum lysate which provides direct proof that the gene cloned encodes hexokinase of the parasite.
Olafsson
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P
Department PRTB, F. Hoffmann-La Roche, Ltd., Basel, Switzerland.
Matile
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H
Certa
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U
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D001483:Base Sequence; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D016054:DNA, Protozoan / Q000235:genetics; D015723:Gene Library; D017125:Genes, Protozoan; D006019:Glycolysis; D006593:Hexokinase / Q000235:genetics* / Q000276:immunology; D008969:Molecular Sequence Data; D010963:Plasmodium falciparum / Q000201:enzymology* / Q000235:genetics* / Q000276:immunology; D012333:RNA, Messenger / Q000235:genetics; D016053:RNA, Protozoan / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology
D000911:Antibodies, Monoclonal; D016054:DNA, Protozoan; D012333:RNA, Messenger; D016053:RNA, Protozoan; D011994:Recombinant Proteins; D006593:Hexokinase