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Import of host δ-aminolevulinate dehydratase into the malarial parasite: Identification of a new drug target

Nature Medicine volume 6, pages 898–903 (2000)Cite this article

Abstract

The parasite Plasmodium berghei imports the enzyme δ-aminolevulinate dehydratase (ALAD), and perhaps the subsequent enzymes of the pathway from the host red blood cell to sustain heme synthesis. Here we have studied the mechanism of this import. A 65-kDa protein on the P. berghei membrane specifically bound to mouse red blood cell ALAD, and a 93-amino-acid fragment (ALAD-ΔNC) of the host erythrocyte ALAD was able to compete with the full-length enzyme for binding to the P. berghei membrane. ALAD-ΔNC was taken up by the infected red blood cell when added to a culture of P. falciparum and this led to a substantial decrease in ALAD protein and enzyme activity and, subsequently, heme synthesis in the parasite, resulting in its death.

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Figure 1: Identification of the protein on P. berghei membrane binding to mouse red blood cell ALAD.
Figure 2: Analysis of binding of mouse red blood cell ALAD deletion mutants to solubilized P. berghei membrane at a pH of 6.5. A nitrocellulose filter binding assay was used5.
Figure 3: Confocal microscopy of the uptake of ALAD-ΔNC and Gal 4 polypeptides added to P. falciparum culture and their effects on endogenous ALAD.
Figure 4: Localization of ALAD-ΔNC, Gal 4 polypeptide and endogenous host red blood cell ALAD in the red blood cell cytoplasm, parasite and food vacuole in P. falciparum culture.
Figure 5: Effect of the addition of ALAD-ΔNC to P. falciparum culture. The purified 93-amino-acid fragment expressed in E. coli was added to P. falciparum culture at a concentration of 5 μM.
Figure 6: The inhibitory effect of ALAD-ΔNC on the import of host red blood cell ALAD into P. falciparum in culture, explained by a model is based on the competition between ALAD-ΔNC and ALAD in the food vacuole to bind to the 65-kDa binding protein present on the food vacuolar membrane.

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Acknowledgements

This work was supported by grants from the Department of Biotechnology and the Council of Scientific & Industrial Research (New Delhi, India).

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  1. Z.Q. Bonday and S. Dhanasekaran: Z.Q.B. and S.D. contributed equally to this study.

Authors and Affiliations

  1. Department of Biochemistry, Indian Institute of Science, Bangalore, 560 012, India

    Z.Q. Bonday, S. Dhanasekaran, P.N. Rangarajan & G. Padmanaban

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  1. Z.Q. Bonday
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  2. S. Dhanasekaran
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  3. P.N. Rangarajan
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  4. G. Padmanaban
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Correspondence to G. Padmanaban.

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Bonday, Z., Dhanasekaran, S., Rangarajan, P. et al. Import of host δ-aminolevulinate dehydratase into the malarial parasite: Identification of a new drug target. Nat Med 6, 898–903 (2000). https://doi.org/10.1038/78659

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