Artemisinin-resistant Plasmodium falciparum Kelch13 mutant proteins display reduced heme-binding affinity and decreased artemisinin activation

Abstract The potency of frontline antimalarial drug artemisinin (ART) derivatives is triggered by heme-induced cleavage of the endoperoxide bond to form reactive heme-ART alkoxy radicals and covalent heme-ART adducts, which are highly toxic to the parasite. ART-resistant (ART-R) parasites with mutat...

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Main Authors: Abdur Rahman, Sabahat Tamseel, Smritikana Dutta, Nawaal Khan, Mohammad Faaiz, Harshita Rastogi, Jyoti Rani Nath, Kasturi Haldar, Pramit Chowdhury, Ashish, Souvik Bhattacharjee
Format: Article
Language:English
Published: Nature Portfolio 2024-11-01
Series:Communications Biology
Online Access:https://doi.org/10.1038/s42003-024-07178-2
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author Abdur Rahman
Sabahat Tamseel
Smritikana Dutta
Nawaal Khan
Mohammad Faaiz
Harshita Rastogi
Jyoti Rani Nath
Kasturi Haldar
Pramit Chowdhury
Ashish
Souvik Bhattacharjee
author_facet Abdur Rahman
Sabahat Tamseel
Smritikana Dutta
Nawaal Khan
Mohammad Faaiz
Harshita Rastogi
Jyoti Rani Nath
Kasturi Haldar
Pramit Chowdhury
Ashish
Souvik Bhattacharjee
author_sort Abdur Rahman
collection DOAJ
description Abstract The potency of frontline antimalarial drug artemisinin (ART) derivatives is triggered by heme-induced cleavage of the endoperoxide bond to form reactive heme-ART alkoxy radicals and covalent heme-ART adducts, which are highly toxic to the parasite. ART-resistant (ART-R) parasites with mutations in the Plasmodium falciparum Kelch-containing protein Kelch13 (PfKekch13) exhibit impaired hemoglobin uptake, reduced yield of hemoglobin-derived heme, and thus decreased ART activation. However, any direct involvement of PfKelch13 in heme-mediated ART activation has not been reported. Here, we show that the purified recombinant PfKelch13 wild-type (WT) protein displays measurable binding affinity for iron and heme, the main effectors for ART activation. The heme-binding property is also exhibited by the native PfKelch13 protein from parasite culture. The two ART-R recombinant PfKelch13 mutants (C580Y and R539T) display weaker heme binding affinities compared to the ART-sensitive WT and A578S mutant proteins, which further translates into reduced yield of heme-ART derivatives when ART is incubated with the heme molecules bound to the mutant PfKelch13 proteins. In conclusion, this study provides the first evidence for ART activation via the heme-binding propensity of PfKelch13. This mechanism may contribute to the modulation of ART-R levels in malaria parasites through a novel function of PfKelch13.
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spelling doaj-art-7ae26975befc4a84b88c8d9c651d521e2024-11-17T12:42:47ZengNature PortfolioCommunications Biology2399-36422024-11-017112310.1038/s42003-024-07178-2Artemisinin-resistant Plasmodium falciparum Kelch13 mutant proteins display reduced heme-binding affinity and decreased artemisinin activationAbdur Rahman0Sabahat Tamseel1Smritikana Dutta2Nawaal Khan3Mohammad Faaiz4Harshita Rastogi5Jyoti Rani Nath6Kasturi Haldar7Pramit Chowdhury8Ashish9Souvik Bhattacharjee10Special Centre for Molecular Medicine, Jawaharlal Nehru UniversitySpecial Centre for Molecular Medicine, Jawaharlal Nehru UniversitySpecial Centre for Molecular Medicine, Jawaharlal Nehru UniversitySpecial Centre for Molecular Medicine, Jawaharlal Nehru UniversitySpecial Centre for Molecular Medicine, Jawaharlal Nehru UniversityDepartment of Chemistry, Indian Institute of Technology (IIT) DelhiDepartment of Chemistry, Indian Institute of Technology (IIT) DelhiDepartment of Biological Sciences, University of Notre Dame, Notre DameDepartment of Chemistry, Indian Institute of Technology (IIT) DelhiCouncil of Scientific and Industrial Research-Institute of Microbial TechnologySpecial Centre for Molecular Medicine, Jawaharlal Nehru UniversityAbstract The potency of frontline antimalarial drug artemisinin (ART) derivatives is triggered by heme-induced cleavage of the endoperoxide bond to form reactive heme-ART alkoxy radicals and covalent heme-ART adducts, which are highly toxic to the parasite. ART-resistant (ART-R) parasites with mutations in the Plasmodium falciparum Kelch-containing protein Kelch13 (PfKekch13) exhibit impaired hemoglobin uptake, reduced yield of hemoglobin-derived heme, and thus decreased ART activation. However, any direct involvement of PfKelch13 in heme-mediated ART activation has not been reported. Here, we show that the purified recombinant PfKelch13 wild-type (WT) protein displays measurable binding affinity for iron and heme, the main effectors for ART activation. The heme-binding property is also exhibited by the native PfKelch13 protein from parasite culture. The two ART-R recombinant PfKelch13 mutants (C580Y and R539T) display weaker heme binding affinities compared to the ART-sensitive WT and A578S mutant proteins, which further translates into reduced yield of heme-ART derivatives when ART is incubated with the heme molecules bound to the mutant PfKelch13 proteins. In conclusion, this study provides the first evidence for ART activation via the heme-binding propensity of PfKelch13. This mechanism may contribute to the modulation of ART-R levels in malaria parasites through a novel function of PfKelch13.https://doi.org/10.1038/s42003-024-07178-2
spellingShingle Abdur Rahman
Sabahat Tamseel
Smritikana Dutta
Nawaal Khan
Mohammad Faaiz
Harshita Rastogi
Jyoti Rani Nath
Kasturi Haldar
Pramit Chowdhury
Ashish
Souvik Bhattacharjee
Artemisinin-resistant Plasmodium falciparum Kelch13 mutant proteins display reduced heme-binding affinity and decreased artemisinin activation
Communications Biology
title Artemisinin-resistant Plasmodium falciparum Kelch13 mutant proteins display reduced heme-binding affinity and decreased artemisinin activation
title_full Artemisinin-resistant Plasmodium falciparum Kelch13 mutant proteins display reduced heme-binding affinity and decreased artemisinin activation
title_fullStr Artemisinin-resistant Plasmodium falciparum Kelch13 mutant proteins display reduced heme-binding affinity and decreased artemisinin activation
title_full_unstemmed Artemisinin-resistant Plasmodium falciparum Kelch13 mutant proteins display reduced heme-binding affinity and decreased artemisinin activation
title_short Artemisinin-resistant Plasmodium falciparum Kelch13 mutant proteins display reduced heme-binding affinity and decreased artemisinin activation
title_sort artemisinin resistant plasmodium falciparum kelch13 mutant proteins display reduced heme binding affinity and decreased artemisinin activation
url https://doi.org/10.1038/s42003-024-07178-2
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