Abstract
Parasites of the phylum Apicomplexa cause substantial morbidity, mortality and economic losses, and new medicines to treat them are needed urgently1,2. The shikimate pathway is an attractive target for herbicides and antimicrobial agents because it is essential in algae, higher plants, bacteria and fungi, but absent from mammals3,4. Here we present biochemical, genetic and chemotherapeutic evidence for the presence of enzymes of the shikimate pathway in apicomplexan parasites. In vitro growth of Toxoplasma gondii, Plasmodium falciparum (malaria) and Cryptosporidium parvum was inhibited by the herbicide glyphosate, a well-characterized inhibitor3 of the shikimate pathway enzyme 5-enolpyruvyl shikimate 3-phosphate synthase. This effect on T. gondii and P. falciparum was reversed by treatment with p-aminobenzoate, which suggests that the shikimate pathway supplies folate precursors for their growth. Glyphosate in combination with pyrimethamine limited T. gondii infection in mice. Four shikimate pathway enzymes were detected in extracts of T. gondii and glyphosate inhibited 5-enolpyruvyl shikimate 3-phosphate synthase activity. Genes encoding chorismate synthase, the final shikimate pathway enzyme, were cloned from T. gondii and P. falciparum. This discovery of a functional shikimate pathway in apicomplexan parasites provides several targets for the development of new antiparasite agents.
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Acknowledgements
We thank L. Mets for information about plants; D. Sibley and the Toxoplasma EST Project at Washington University for the putative chorismate synthase EST; M. Gottlieb, J. Boothroyd, K. Kim, L. Weiss, A. Lapthorn, A. Martin, M. Rust and U. Gross for discussions; and V. Aitchison and E. Holfels for assistance in preparation of this manuscript. F.R. was a recipient of a Royal Pathological Society of Great Britain and Northern Ireland Travelling Fellowship, C.W.R. was a Fulbright Scholar, and a Michael Reese Physicians' Research and Education Foundation Postdoctoral Fellow and Visiting Scholar and is a Glaxo Jack Lecturer. R.M. is the Jules and Doris Stein Research to Prevent Blindness Professor at The University of Chicago. This work was supported by the NIH NIAID TMP program, the Toxoplasmosis Research Institute, the Research to Prevent Blindness Foundation, an NIH contract from NIH NIAID (the AIDS Opportunistic Infection Drug Discovery), and the WHO-NIH-WRAIR Drug Development Programs, The Wellcome Trust and the European Union.
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Roberts, F., Roberts, C., Johnson, J. et al. Evidence for the shikimate pathway in apicomplexan parasites. Nature 393, 801–805 (1998). https://doi.org/10.1038/31723
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DOI: https://doi.org/10.1038/31723
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