Related Experiment Video
Updated: Jun 18, 2025
Author Spotlight: Identifying Compensatory Pathways in Malaria Parasites Containing Hypomorphic Allele of Essential Protein Kinases
Published on: November 22, 2024
Mutations in Plasmodium falciparum Kelch13 (PfK13) dysregulates PI3K/AKT signalling pathway in artemisinin resistance
Kanika Verma1, Yashika Singh1, Balu Kamaraj2
1Department of Molecular Epidemiology, ICMR-NIMR, Dwarka, Delhi, 110077, India.
Insights
Mutations in the PfK13 protein destabilize its structure, impacting PfPI3K binding and contributing to artemisinin resistance in Plasmodium falciparum. Further research is needed to understand this interaction and combat resistance.
Area of Science:
- Molecular Biology
- Parasitology
- Drug Resistance
Background:
- Mutations in the Plasmodium falciparum Kelch 13 (PfK13) protein are linked to artemisinin resistance.
- PfK13 protein regulates ubiquitination and the PI3K/AKT pathway.
- Mutant PfK13 variations can impair substrate binding, increasing PfPI3K levels.
Purpose of the Study:
- To investigate the impact of PfK13 mutations on protein stability and PfPI3K binding.
- To understand the molecular mechanisms underlying artemisinin resistance in Plasmodium falciparum.
Main Methods:
- Utilized computational tools (DUET, DynaMut2, mCSM, iStable 2.0, I-Mutant 2.0, MuPro) to assess protein stability.
- Employed HADDOCK 2.4 for molecular docking to analyze binding between wild-type and mutant PfK13 with PfPI3K.
- Validated binding affinity using MM_GBSA to calculate binding free energy (DDG).
Main Results:
- Seven out of nine artemisinin resistance-associated PfK13 mutations decreased protein stability.
- HADDOCK scores indicated significant alterations in PfK13-PfPI3K binding affinity due to mutations.
- Analysis revealed a loss of interactions between mutant PfK13 and PfPI3K, evidenced by binding free energy changes.
Conclusions:
- PfK13 mutations destabilize the protein structure and disrupt PfPI3K binding.
- These findings provide a basis for further research into the PfK13-PfPI3K interaction to overcome artemisinin resistance.
- Further validation is required to confirm these preliminary results.
Background:
PfK13 protein mutations are associated with the emergence of artemisinin resistance in Plasmodium falciparum. PfK13 protein is essential for mediating ubiquitination and controlling the PI3K/AKT pathway. Mutant PfK13 variations can interfere with substrate binding, especially with PfPI3K, which raises PfPI3K levels.
Methods:
DUET, DynaMut2, mCSM, iStable 2.0, I-Mutant 2.0, and MuPro were utilized to study the protein stability and protein-substrate binding was studied using HADDOCK 2.4 docking algorithm between Wild-type and mutant PfK13 with the helical and catalytic domain of PfPI3K.
Results:
i-Stable server analysis predicted that seven, out of the nine mutations associated with artemisinin resistance (F446I, Y493H, R539T, I543T, P553L, R561H, C580Y) reduced the protein stability. HADDOCK scores of the catalytic domain underscores the significant impact of the reported mutations on the binding affinity of the PfK13 protein. Further validation through the MM_GBSA technique, the binding free energy (DDG) between the wild-type and the mutant PfK13 protein analysis revealed a loss of interactions resulting from mutations in PfK13.
Conclusion:
The study finding suggest that mutations in the PfK13 cause destabilization in the protein structure and affects the binding of PfPI3K. Although the findings remain preliminary and require further validation, it provides the basis for further research considering the importance of the interaction of PfK13 and PfPI3K to overcome the impact of artemisinin resistance.
