Identification and structural validation of purine nucleoside phosphorylase from Plasmodium falciparum as a target of MMV000848

About 247 million cases of malaria occurred in 2021 with Plasmodium falciparum accounting for the majority of 619,000 deaths. In the absence of a widely available vaccine, chemotherapy remains crucial to prevent, treat, and contain the disease. The efficacy of several drugs currently used in the cli...

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Main Authors: Chung, Zara, Lin, Jianqing, Wirjanata, Grennady, Dziekan, Jerzy Michal, El Sahili, Abbas, Preiser, Peter Rainer, Bozdech, Zbynek, Lescar, Julien
Other Authors: School of Biological Sciences
Format: Article
Language:English
Published: 2024
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Online Access:https://hdl.handle.net/10356/174722
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Institution: Nanyang Technological University
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spelling sg-ntu-dr.10356-1747222024-04-08T15:32:25Z Identification and structural validation of purine nucleoside phosphorylase from Plasmodium falciparum as a target of MMV000848 Chung, Zara Lin, Jianqing Wirjanata, Grennady Dziekan, Jerzy Michal El Sahili, Abbas Preiser, Peter Rainer Bozdech, Zbynek Lescar, Julien School of Biological Sciences NTU Institute of Structural Biology Medicine, Health and Life Sciences Malaria Enzyme inhibition About 247 million cases of malaria occurred in 2021 with Plasmodium falciparum accounting for the majority of 619,000 deaths. In the absence of a widely available vaccine, chemotherapy remains crucial to prevent, treat, and contain the disease. The efficacy of several drugs currently used in the clinic is likely to suffer from the emergence of resistant parasites. A global effort to identify lead compounds led to several initiatives such as the Medicine for Malaria Ventures (MMV), a repository of compounds showing promising efficacy in killing the parasite in cell-based assays. Here, we used mass spectrometry coupled with cellular thermal shift assay to identify putative protein targets of MMV000848, a compound with an in vitro EC50 of 0.5 μM against the parasite. Thermal shift assays showed a strong increase of P. falciparum purine nucleoside phosphorylase (PfPNP) melting temperature by up to 15 °C upon incubation with MMV000848. Binding and enzymatic assays returned a KD of 1.52 ± 0.495 μM and an IC50 value of 21.5 ± 2.36 μM. The inhibition is competitive with respect to the substrate, as confirmed by a cocrystal structure of PfPNP bound with MMV000848 at the active site, determined at 1.85 Å resolution. In contrast to transition states inhibitors, MMV000848 specifically inhibits the parasite enzyme but not the human ortholog. An isobologram analysis shows subadditivity with immucillin H and with quinine respectively, suggesting overlapping modes of action between these compounds. These results point to PfPNP as a promising antimalarial target and suggest avenues to improve inhibitor potency. National Research Foundation (NRF) Published version This work was supported by grant NRF-CRP24-2020-005 to J. L., P. R. P., and Z. B. 2024-04-08T06:29:36Z 2024-04-08T06:29:36Z 2024 Journal Article Chung, Z., Lin, J., Wirjanata, G., Dziekan, J. M., El Sahili, A., Preiser, P. R., Bozdech, Z. & Lescar, J. (2024). Identification and structural validation of purine nucleoside phosphorylase from Plasmodium falciparum as a target of MMV000848. Journal of Biological Chemistry, 300(1), 105586-. https://dx.doi.org/10.1016/j.jbc.2023.105586 0021-9258 https://hdl.handle.net/10356/174722 10.1016/j.jbc.2023.105586 38141766 2-s2.0-85182280374 1 300 105586 en NRF-CRP24-2020-005 Journal of Biological Chemistry © 2023 The Authors. Published by Elsevier Inc on behalf of American Society for Biochemistry and Molecular Biology. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). application/pdf
institution Nanyang Technological University
building NTU Library
continent Asia
country Singapore
Singapore
content_provider NTU Library
collection DR-NTU
language English
topic Medicine, Health and Life Sciences
Malaria
Enzyme inhibition
spellingShingle Medicine, Health and Life Sciences
Malaria
Enzyme inhibition
Chung, Zara
Lin, Jianqing
Wirjanata, Grennady
Dziekan, Jerzy Michal
El Sahili, Abbas
Preiser, Peter Rainer
Bozdech, Zbynek
Lescar, Julien
Identification and structural validation of purine nucleoside phosphorylase from Plasmodium falciparum as a target of MMV000848
description About 247 million cases of malaria occurred in 2021 with Plasmodium falciparum accounting for the majority of 619,000 deaths. In the absence of a widely available vaccine, chemotherapy remains crucial to prevent, treat, and contain the disease. The efficacy of several drugs currently used in the clinic is likely to suffer from the emergence of resistant parasites. A global effort to identify lead compounds led to several initiatives such as the Medicine for Malaria Ventures (MMV), a repository of compounds showing promising efficacy in killing the parasite in cell-based assays. Here, we used mass spectrometry coupled with cellular thermal shift assay to identify putative protein targets of MMV000848, a compound with an in vitro EC50 of 0.5 μM against the parasite. Thermal shift assays showed a strong increase of P. falciparum purine nucleoside phosphorylase (PfPNP) melting temperature by up to 15 °C upon incubation with MMV000848. Binding and enzymatic assays returned a KD of 1.52 ± 0.495 μM and an IC50 value of 21.5 ± 2.36 μM. The inhibition is competitive with respect to the substrate, as confirmed by a cocrystal structure of PfPNP bound with MMV000848 at the active site, determined at 1.85 Å resolution. In contrast to transition states inhibitors, MMV000848 specifically inhibits the parasite enzyme but not the human ortholog. An isobologram analysis shows subadditivity with immucillin H and with quinine respectively, suggesting overlapping modes of action between these compounds. These results point to PfPNP as a promising antimalarial target and suggest avenues to improve inhibitor potency.
author2 School of Biological Sciences
author_facet School of Biological Sciences
Chung, Zara
Lin, Jianqing
Wirjanata, Grennady
Dziekan, Jerzy Michal
El Sahili, Abbas
Preiser, Peter Rainer
Bozdech, Zbynek
Lescar, Julien
format Article
author Chung, Zara
Lin, Jianqing
Wirjanata, Grennady
Dziekan, Jerzy Michal
El Sahili, Abbas
Preiser, Peter Rainer
Bozdech, Zbynek
Lescar, Julien
author_sort Chung, Zara
title Identification and structural validation of purine nucleoside phosphorylase from Plasmodium falciparum as a target of MMV000848
title_short Identification and structural validation of purine nucleoside phosphorylase from Plasmodium falciparum as a target of MMV000848
title_full Identification and structural validation of purine nucleoside phosphorylase from Plasmodium falciparum as a target of MMV000848
title_fullStr Identification and structural validation of purine nucleoside phosphorylase from Plasmodium falciparum as a target of MMV000848
title_full_unstemmed Identification and structural validation of purine nucleoside phosphorylase from Plasmodium falciparum as a target of MMV000848
title_sort identification and structural validation of purine nucleoside phosphorylase from plasmodium falciparum as a target of mmv000848
publishDate 2024
url https://hdl.handle.net/10356/174722
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