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Insights into antifolate resistance from malarial DHFR-TS structures

Nature Structural & Molecular Biology volume 10, pages 357–365 (2003)Cite this article

Abstract

Plasmodium falciparum dihydrofolate reductase–thymidylate synthase (PfDHFR-TS) is an important target of antimalarial drugs. The efficacy of this class of DHFR-inhibitor drugs is now compromised because of mutations that prevent drug binding yet retain enzyme activity. The crystal structures of PfDHFR-TS from the wild type (TM4/8.2) and the quadruple drug-resistant mutant (V1/S) strains, in complex with a potent inhibitor WR99210, as well as the resistant double mutant (K1 CB1) with the antimalarial pyrimethamine, reveal features for overcoming resistance. In contrast to pyrimethamine, the flexible side chain of WR99210 can adopt a conformation that fits well in the active site, thereby contributing to binding. The single-chain bifunctional PfDHFR-TS has a helical insert between the DHFR and TS domains that is involved in dimerization and domain organization. Moreover, positively charged grooves on the surface of the dimer suggest a function in channeling of substrate from TS to DHFR active sites. These features provide possible approaches for the design of new drugs to overcome antifolate resistance.

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Figure 1: Homology and structure-based alignment of DHFR and junction region from various organisms.
Figure 2: Overview of the wild type PfDHFR-TS structure.
Figure 3: Inhibitors WR99210 and pyrimethamine bound at the DHFR active site.
Figure 4: Enzyme–inhibitor interactions at the active site.
Figure 5: Surface electrostatic potential on the wild type PfDHFR-TS dimer.

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Acknowledgements

We thank the Wellcome Trust for the Collaborative Research Grant to Y.Y. and M.W. The present work was also partly supported by grants from the EU, Medicines for Malaria Venture (MMV), the Special Programme for Research and Training in Tropical Diseases (TDR)/United Nations Development Programme/the World Bank/the World Health Organization and Thailand Tropical Diseases Research (T2) Programmes to Y.Y. and S.K. and from the TDR to W.S. We are grateful to CCLRC, ESRF, EMBL and NSLS for use of synchrotron facilities and to W.N. Lipscomb for his suggestions and comments on the manuscript. We also thank S. Thaithong, Department of Biology, Faculty of Science, Chulalongkorn University (TM4/8.2 and K1 CB1), and D. Kyle through MR4 (V1/S) for sources of the parasite strains.

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  1. Department of Biochemistry, Faculty of Science, Mahidol University, Rama 6 Road, Bangkok, 10400, Thailand

    Jirundon Yuvaniyama & Worachart Sirawaraporn

  2. BIOTEC, National Science and Technology Development Agency, Science Park, 113 Phaholyothin Road, Klong 1, Klong Luang, 12120, Pathumthani, Thailand

    Penchit Chitnumsub, Sumalee Kamchonwongpaisan, Jarunee Vanichtanankul & Yongyuth Yuthavong

  3. Institute of Cell and Molecular Biology, The University of Edinburgh, Kings Buildings, Mayfield Road, Edinburgh, EH9 3JR, UK

    Paul Taylor & Malcolm D. Walkinshaw

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Yuvaniyama, J., Chitnumsub, P., Kamchonwongpaisan, S. et al. Insights into antifolate resistance from malarial DHFR-TS structures. Nat Struct Mol Biol 10, 357–365 (2003). https://doi.org/10.1038/nsb921

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