Abstract：
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.
CONCLUSIONS: 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.
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.
CONCLUSIONS: 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.
摘要：
背景：PfK13蛋白突变与恶性疟原虫青蒿素抗性的出现有关。PfK13蛋白对于介导泛素化和控制PI3K/AKT途径至关重要。突变体PfK13变异会干扰底物结合，尤其是PfPI3K,这提高了PfPI3K水平。
方法：DUET，DynaMut2,mCSM,iStable2.0，I-Mutant2.0和MuPro用于研究蛋白质稳定性，并使用HADDOCK2.4对接算法研究了野生型和突变型PfK13与PfPI3K的螺旋和催化结构域之间的蛋白质-底物结合。
结果：i-Stable服务器分析预测，在与青蒿素抗性相关的9个突变中(F446I，Y493H,R539T,I543T,P553L,R561H,C580Y）降低了蛋白质的稳定性。催化结构域的HADDOCK评分强调了所报道的突变对PfK13蛋白的结合亲和力的显著影响。通过MM_GBSA技术进一步验证，野生型和突变型PfK13蛋白之间的结合自由能(ddG)分析揭示了由PfK13突变引起的相互作用的丧失。
结论：研究发现表明，PfK13中的突变导致蛋白质结构不稳定，并影响PfPI3K的结合。尽管调查结果仍是初步的，需要进一步验证，考虑到PfK13和PfPI3K相互作用对克服青蒿素耐药性影响的重要性，为进一步研究提供了基础。
方法：DUET，DynaMut2,mCSM,iStable2.0，I-Mutant2.0和MuPro用于研究蛋白质稳定性，并使用HADDOCK2.4对接算法研究了野生型和突变型PfK13与PfPI3K的螺旋和催化结构域之间的蛋白质-底物结合。
结果：i-Stable服务器分析预测，在与青蒿素抗性相关的9个突变中(F446I，Y493H,R539T,I543T,P553L,R561H,C580Y）降低了蛋白质的稳定性。催化结构域的HADDOCK评分强调了所报道的突变对PfK13蛋白的结合亲和力的显著影响。通过MM_GBSA技术进一步验证，野生型和突变型PfK13蛋白之间的结合自由能(ddG)分析揭示了由PfK13突变引起的相互作用的丧失。
结论：研究发现表明，PfK13中的突变导致蛋白质结构不稳定，并影响PfPI3K的结合。尽管调查结果仍是初步的，需要进一步验证，考虑到PfK13和PfPI3K相互作用对克服青蒿素耐药性影响的重要性，为进一步研究提供了基础。
