Assessing the efficacy of iso-mukaadial acetate and betulinic acid against selected Plasmodium falciparum glycolytic pathway proteins: in silico and in vitro studies
Abstract Malaria is the extensive health concern in sub-Saharan Africa, with Plasmodium falciparum being the most lethal strain. The continued emergence of drug-resistant P. falciparum advocates for the development of new antimalarials. Our current study aimed to effectively explore the interaction...
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2025-01-01
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| Online Access: | https://doi.org/10.1186/s13065-025-01380-x |
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| author | Lindiwe Khumbuzile Zuma Ofentse Jacob Pooe Nonduduzo Hlengiwe Mabaso John Alake Vincent A. Obakachi Sinethemba Yakobi Nothando Gasa Rajshekhar Karpoormath Mthokozisi Simelane |
| author_facet | Lindiwe Khumbuzile Zuma Ofentse Jacob Pooe Nonduduzo Hlengiwe Mabaso John Alake Vincent A. Obakachi Sinethemba Yakobi Nothando Gasa Rajshekhar Karpoormath Mthokozisi Simelane |
| author_sort | Lindiwe Khumbuzile Zuma |
| collection | DOAJ |
| description | Abstract Malaria is the extensive health concern in sub-Saharan Africa, with Plasmodium falciparum being the most lethal strain. The continued emergence of drug-resistant P. falciparum advocates for the development of new antimalarials. Our current study aimed to effectively explore the interaction capabilities of iso-mukaadial acetate (IMA) and betulinic acid (BA) against two essential P. falciparum glycolytic pathway proteins, PfLDH and PfHk. Recombinant PfLDH and PfHk were independently expressed in E. coli BL21 (DE3) cells and subsequently purified using affinity chromatography. Protein–ligand interaction studies probed in silico and in vitro approaches. Parasite inhibition studies confirmed potent antimalarial activity against the P. falciparum NF54 strains, with BA and IMA showing IC50 values of 1.27 µg/ml and 1.03 µg/ml against the asexual stage of P. falciparum, respectively. FTIR experiments confirmed interactions between the compounds and the secondary structure of the proteins. Direct protein–ligand interaction studies analysis using microscale thermophoresis (MST) showed a KD value of 0.1036 ± 0.6001 µM for the PfLDH-BA complex and 0.7473 ± 0.3554 µM KD value for PfLDH-IMA. Meanwhile, PfHk-IMA had 0.39701 ± 0.16298 µM KD value, while the PfHk-BA complex had no interaction detected. Molecular docking and molecular dynamics simulation studies were used to measure and confirm the interactive strength of complexes. Molecular docking reported a binding score of − 1.155 kcal/mol for the PfLDH-BA complex and a binding score of − 3.200 kcal/mol for PfLDH-IMA. The PfHk-BA complex had − 2.871 kcal/mol and PfHk-IMA complex had − 4.225 kcal/mol binding score. In conclusion, BA and IMA compounds had better interactions and remained bound within the binding sites of the glycolytic pathway proteins (PfLDH and PfHk). |
| format | Article |
| id | doaj-art-c68f3f77abfc4bec92dbf6101e640342 |
| institution | Kabale University |
| issn | 2661-801X |
| language | English |
| publishDate | 2025-01-01 |
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| series | BMC Chemistry |
| spelling | doaj-art-c68f3f77abfc4bec92dbf6101e6403422025-01-19T12:08:32ZengBMCBMC Chemistry2661-801X2025-01-0119111310.1186/s13065-025-01380-xAssessing the efficacy of iso-mukaadial acetate and betulinic acid against selected Plasmodium falciparum glycolytic pathway proteins: in silico and in vitro studiesLindiwe Khumbuzile Zuma0Ofentse Jacob Pooe1Nonduduzo Hlengiwe Mabaso2John Alake3Vincent A. Obakachi4Sinethemba Yakobi5Nothando Gasa6Rajshekhar Karpoormath7Mthokozisi Simelane8Discipline of Biochemistry, School of Life Sciences, University of KwaZulu-NatalDiscipline of Biochemistry, School of Life Sciences, University of KwaZulu-NatalDiscipline of Biochemistry, School of Life Sciences, University of KwaZulu-NatalDepartment of Pharmaceutical Chemistry, Discipline of Pharmaceutical Sciences, College of Health Sciences, University of KwaZulu-NatalDepartment of Pharmaceutical Chemistry, Discipline of Pharmaceutical Sciences, College of Health Sciences, University of KwaZulu-NatalDiscipline of Biochemistry, School of Life Sciences, University of KwaZulu-NatalDiscipline of Biochemistry, School of Life Sciences, University of KwaZulu-NatalDepartment of Pharmaceutical Chemistry, Discipline of Pharmaceutical Sciences, College of Health Sciences, University of KwaZulu-NatalDepartment of Biochemistry, University of JohannesburgAbstract Malaria is the extensive health concern in sub-Saharan Africa, with Plasmodium falciparum being the most lethal strain. The continued emergence of drug-resistant P. falciparum advocates for the development of new antimalarials. Our current study aimed to effectively explore the interaction capabilities of iso-mukaadial acetate (IMA) and betulinic acid (BA) against two essential P. falciparum glycolytic pathway proteins, PfLDH and PfHk. Recombinant PfLDH and PfHk were independently expressed in E. coli BL21 (DE3) cells and subsequently purified using affinity chromatography. Protein–ligand interaction studies probed in silico and in vitro approaches. Parasite inhibition studies confirmed potent antimalarial activity against the P. falciparum NF54 strains, with BA and IMA showing IC50 values of 1.27 µg/ml and 1.03 µg/ml against the asexual stage of P. falciparum, respectively. FTIR experiments confirmed interactions between the compounds and the secondary structure of the proteins. Direct protein–ligand interaction studies analysis using microscale thermophoresis (MST) showed a KD value of 0.1036 ± 0.6001 µM for the PfLDH-BA complex and 0.7473 ± 0.3554 µM KD value for PfLDH-IMA. Meanwhile, PfHk-IMA had 0.39701 ± 0.16298 µM KD value, while the PfHk-BA complex had no interaction detected. Molecular docking and molecular dynamics simulation studies were used to measure and confirm the interactive strength of complexes. Molecular docking reported a binding score of − 1.155 kcal/mol for the PfLDH-BA complex and a binding score of − 3.200 kcal/mol for PfLDH-IMA. The PfHk-BA complex had − 2.871 kcal/mol and PfHk-IMA complex had − 4.225 kcal/mol binding score. In conclusion, BA and IMA compounds had better interactions and remained bound within the binding sites of the glycolytic pathway proteins (PfLDH and PfHk).https://doi.org/10.1186/s13065-025-01380-xMalariaPlasmodium falciparumMedicinal compoundsInteraction studies |
| spellingShingle | Lindiwe Khumbuzile Zuma Ofentse Jacob Pooe Nonduduzo Hlengiwe Mabaso John Alake Vincent A. Obakachi Sinethemba Yakobi Nothando Gasa Rajshekhar Karpoormath Mthokozisi Simelane Assessing the efficacy of iso-mukaadial acetate and betulinic acid against selected Plasmodium falciparum glycolytic pathway proteins: in silico and in vitro studies BMC Chemistry Malaria Plasmodium falciparum Medicinal compounds Interaction studies |
| title | Assessing the efficacy of iso-mukaadial acetate and betulinic acid against selected Plasmodium falciparum glycolytic pathway proteins: in silico and in vitro studies |
| title_full | Assessing the efficacy of iso-mukaadial acetate and betulinic acid against selected Plasmodium falciparum glycolytic pathway proteins: in silico and in vitro studies |
| title_fullStr | Assessing the efficacy of iso-mukaadial acetate and betulinic acid against selected Plasmodium falciparum glycolytic pathway proteins: in silico and in vitro studies |
| title_full_unstemmed | Assessing the efficacy of iso-mukaadial acetate and betulinic acid against selected Plasmodium falciparum glycolytic pathway proteins: in silico and in vitro studies |
| title_short | Assessing the efficacy of iso-mukaadial acetate and betulinic acid against selected Plasmodium falciparum glycolytic pathway proteins: in silico and in vitro studies |
| title_sort | assessing the efficacy of iso mukaadial acetate and betulinic acid against selected plasmodium falciparum glycolytic pathway proteins in silico and in vitro studies |
| topic | Malaria Plasmodium falciparum Medicinal compounds Interaction studies |
| url | https://doi.org/10.1186/s13065-025-01380-x |
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