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Molecular surveillance of Kelch-13 gene in Plasmodium falciparum field isolates from Mayurbhanj District, Odisha, India, and in silico artemisinin-Kelch-13 protein interaction study

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Abstract

The global malaria control and elimination program faces major threats due to the emergence and transmission of the anti-malarial drug-resistant strain of Plasmodium falciparum. Monitoring of artemisinin (ART) resistance marker Kelch-13 in the malaria-endemic region is essential in mitigating the disease’s morbidity and mortality. The current study aimed to generate baseline information for further surveillance in the future. The current research was designed and conducted from July 2019 to June 2021 to monitor Pfkelch13 mutation at the molecular level in the eastern region of India. We also conducted an in silico study to understand the drug-protein interactions between ART and the protein crystal of PfKelch13 (KELCH) with PDB id:4ZGC. The kelch-13 gene was amplified by nested polymerase chain reaction (PCR) and sequenced through the Sanger sequencing method. Reference 3D7 clone (PF3D7_1343700) was used to align and probe all the sequences. The sequence analysis showed the absence of validated or associated mutation in the Kelch-13 propeller domain. The absence of natural selection in drug resistance was confirmed by the Tajima test. Further, in silico interaction studies between the drug ART and the Kelch propeller domain of P. falciparum were evaluated by structure predictions, molecular docking, molecular dynamics (MD) simulations, and estimations of binding free energies for the KELCH-ART complex. The results were compared with the apoprotein (KELCH-APO). The study confirmed the favorable binding of ART with the Kelch-13 propeller domain.

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Acknowledgements

The authors thank Head, P.G. Dept. of Zoology of Berhampur University, Berhampur, for providing the necessary facilities.

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Authors and Affiliations

  1. P.G. Department of Zoology, Berhampur University, Berhampur, Odisha, 760007, India

    Laxman Kumar Murmu & Tapan Kumar Barik

  2. Computational Biology & Bioinformatics Laboratory, P.G. Dept. of Botany, Berhampur University, Berhampur, Odisha, 760007, India

    Madhusmita Panda & Biswa Ranjan Meher

  3. Multi-Disciplinary Research Unit, M.K.C.G Medical College, Berhampur, Odisha, India

    Prasant Purohit

  4. Department of Pharmacology, M.K.C.G Medical College, Berhampur, Odisha, India

    Jayantiprava Behera

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Contributions

Laxman Kumar Murmu: conceptualization, methodology, collection of samples, writing (original draft). Tapan Kumar Barik: conceptualization, supervision, review and editing. Madhusmita Panda and Biswa Ranjan Meher: methodology, writing (original draft). Prasant Purohit: methodology, collection of samples. Jayantiprava Behera: methodology, review.

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Correspondence to Tapan Kumar Barik.

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Murmu, L.K., Panda, M., Meher, B.R. et al. Molecular surveillance of Kelch-13 gene in Plasmodium falciparum field isolates from Mayurbhanj District, Odisha, India, and in silico artemisinin-Kelch-13 protein interaction study. Parasitol Res 122, 717–727 (2023). https://doi.org/10.1007/s00436-023-07784-9

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