Subcellular localization and characterization of chorismate synthase in the apicomplexan Plasmodium falciparum.
The resurgence of drug-resistant apicomplexa, in particular Plasmodium falciparum, the most fatal human malarial parasite, has focused attention on the recent discovery of the shikimate pathway in these organisms, as it may provide the urgently required, novel drug targets resulting from the absence of this pathway in mammals. The direction of a parasiticidal drug design programme obviously requires knowledge of the subcellular localization and indeed full characterization of the possible enzyme targets. Here, we report the cloning and characterization of chorismate synthase from P. falciparum and present the first biochemical and immunological studies of an enzyme of the shikimate pathway from an apicomplexan parasite. We show that this chorismate synthase does not possess an intrinsic flavin reductase activity and is therefore monofunctional like the plant and bacterial chorismate synthases. Highest immunological cross-reactivity was found with a plant chorismate synthase. However, in contrast to the plant enzyme, which is located to the plastid, P. falciparum chorismate synthase is found in the parasite cytosol, akin to the fungal enzymes that possess an intrinsic flavin reductase activity (i.e. are bifunctional). Thus, P. falciparum chorismate synthase has a combination of properties that distinguishes it from other described chorismate synthases.
Fitzpatrick
T
T
ETH-Zürich, Institut für Pflanzenwissenschaften, Universitätstr. 2, CH-8092 Zürich, Switzerland.
Ricken
S
S
Lanzer
M
M
Amrhein
N
N
Macheroux
P
P
Kappes
B
B
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003429:Cross Reactions; D017931:DNA Primers; D008969:Molecular Sequence Data; D019761:Phosphorus-Oxygen Lyases / Q000737:chemistry / Q000378:metabolism*; D010963:Plasmodium falciparum / Q000201:enzymology* / Q000235:genetics; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013347:Subcellular Fractions / Q000201:enzymology*
D017931:DNA Primers; D011994:Recombinant Proteins; C019895:chorismate synthase; D019761:Phosphorus-Oxygen Lyases