Summary of Study ST004425
This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR002796. The data can be accessed directly via it's Project DOI: 10.21228/M87V8Q This work is supported by NIH grant, U2C- DK119886. See: https://www.metabolomicsworkbench.org/about/howtocite.php
| Study ID | ST004425 |
| Study Title | Natural triterpenic phenolic esters target PfA-M17 in Plasmodium falciparum |
| Study Summary | Malaria is a deadly parasitic disease for which innovative treatments are urgently needed. A mixture of eight triterpenic esters (8TTE) was previously identified as important for the antiplasmodial activity of Keetia leucantha twigs, a plant used in traditional medicine in Benin. Thus, 8TTE thus appears as a promising natural product for developing new antimalarial therapeutic strategies against P. falciparum; however, despite the reported in vitro and in vivo activity, the targets of 8TTE are unknown. This study investigated the in vitro mode of action of 8TTE on Plasmodium falciparum parasites in standard culture conditions by a multi-scale integrative study from phenotype to metabolome, including morphologic analysis, enzymatic tests, molecular docking, and metabolomic profiling. This study identified a unique antiplasmodial profile with activity onset in the early-ring stage of the parasite, the inhibition of aminopeptidase PfA-M17 (PlasmoDB PF3D7_1446200), and perturbations in parasite hemoglobin metabolism. Further structure-activity and medicinal chemistry studies are warranted to elaborate on our findings and the potential for 8TTE-related molecules to serve as future antimalarial drugs. |
| Institute | Pennsylvania State University |
| Department | Biochemistry and Molecular Biology |
| Laboratory | Manuel Llinás |
| Last Name | Rangel |
| First Name | Gabriel |
| Address | 491 Pollock Road, University Park, PA, 16802, USA |
| grangel0955@gmail.com | |
| Phone | 8148673527 |
| Submit Date | 2025-12-02 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | mzML |
| Analysis Type Detail | LC-MS |
| Release Date | 2025-12-29 |
| Release Version | 1 |
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Project:
| Project ID: | PR002796 |
| Project DOI: | doi: 10.21228/M87V8Q |
| Project Title: | Natural triterpenic phenolic esters target PfA-M17 in Plasmodium falciparum |
| Project Summary: | Malaria is a deadly parasitic disease for which innovative treatments are urgently needed. A mixture of eight triterpenic esters (8TTE) was previously identified as important for the antiplasmodial activity of Keetia leucantha twigs, a plant used in traditional medicine in Benin. Thus, 8TTE thus appears as a promising natural product for developing new antimalarial therapeutic strategies against P. falciparum; however, despite the reported in vitro and in vivo activity, the targets of 8TTE are unknown. This study investigated the mode of action of 8TTE on Plasmodium falciparum by a multi-scale integrative study from phenotype to metabolome, including phenotypic analysis, enzymatic tests, molecular docking and metabolomic profiling. This study identified a unique antiplasmodial profile with activity onset in the early-ring stage of the parasite, the inhibition of aminopeptidase PfA-M17 (PlasmoDB PF3D7_1446200), and perturbations in parasite hemoglobin metabolism. Further structure-activity and medicinal chemistry studies are warranted to elaborate on our findings and the potential for 8TTE-related molecules to serve as future antimalarial drugs. |
| Institute: | Pennsylvania State University |
| Department: | Biochemistry and Molecular Biology |
| Laboratory: | Manuel Llinás |
| Last Name: | Rangel |
| First Name: | Gabriel |
| Address: | 491 Pollock Road, University Park, PA, 16802, USA |
| Email: | grangel0955@gmail.com |
| Phone: | 8148673527 |
Subject:
| Subject ID: | SU004585 |
| Subject Type: | Cultured cells |
| Subject Species: | Plasmodium falciparum |
| Taxonomy ID: | 5833 |
| Genotype Strain: | 3D7 MR4 |
| Cell Biosource Or Supplier: | MR4 |
| Cell Counts: | 1e6 per sample |
Factors:
Subject type: Cultured cells; Subject species: Plasmodium falciparum (Factor headings shown in green)
| mb_sample_id | local_sample_id | Treatment | Sample source |
|---|---|---|---|
| SA522660 | 4-8-UCL_3 | 8TTE | Plasmodium falciparum 3D7 MR4 |
| SA522661 | 4-8-UCL_2 | 8TTE | Plasmodium falciparum 3D7 MR4 |
| SA522662 | 4-8-UCL_1 | 8TTE | Plasmodium falciparum 3D7 MR4 |
| SA522663 | 4-13-UCL_3 | 8TTE | Plasmodium falciparum 3D7 MR4 |
| SA522664 | 4-13-UCL_2 | 8TTE | Plasmodium falciparum 3D7 MR4 |
| SA522665 | 4-13-UCL_1 | 8TTE | Plasmodium falciparum 3D7 MR4 |
| SA522666 | 4-12-UCL_1 | 8TTE | Plasmodium falciparum 3D7 MR4 |
| SA522667 | 4-12-UCL_2 | 8TTE | Plasmodium falciparum 3D7 MR4 |
| SA522668 | 4-12-UCL_3 | 8TTE | Plasmodium falciparum 3D7 MR4 |
| SA522669 | 4-13-Artemisinin_3 | Artemesinin | Plasmodium falciparum 3D7 MR4 |
| SA522670 | 4-8-Artemisinin_3 | Artemesinin | Plasmodium falciparum 3D7 MR4 |
| SA522671 | 4-8-Artemisinin_2 | Artemesinin | Plasmodium falciparum 3D7 MR4 |
| SA522672 | 4-8-Artemisinin_1 | Artemesinin | Plasmodium falciparum 3D7 MR4 |
| SA522673 | 4-12-Artemisinin_2 | Artemesinin | Plasmodium falciparum 3D7 MR4 |
| SA522674 | 4-12-Artemisinin_1 | Artemesinin | Plasmodium falciparum 3D7 MR4 |
| SA522675 | 4-13-Artemisinin_2 | Artemesinin | Plasmodium falciparum 3D7 MR4 |
| SA522676 | 4-13-Artemisinin_1 | Artemesinin | Plasmodium falciparum 3D7 MR4 |
| SA522677 | 4-12-Artemisinin_3 | Artemesinin | Plasmodium falciparum 3D7 MR4 |
| SA522678 | 4-13-AVQ_1 | Atovaquone | Plasmodium falciparum 3D7 MR4 |
| SA522679 | 4-13-AVQ_2 | Atovaquone | Plasmodium falciparum 3D7 MR4 |
| SA522680 | 4-13-AVQ_3 | Atovaquone | Plasmodium falciparum 3D7 MR4 |
| SA522681 | 4-12-AVQ_1 | Atovaquone | Plasmodium falciparum 3D7 MR4 |
| SA522682 | 4-12-AVQ_2 | Atovaquone | Plasmodium falciparum 3D7 MR4 |
| SA522683 | 4-8-AVQ_3 | Atovaquone | Plasmodium falciparum 3D7 MR4 |
| SA522684 | 4-8-AVQ_2 | Atovaquone | Plasmodium falciparum 3D7 MR4 |
| SA522685 | 4-8-AVQ_1 | Atovaquone | Plasmodium falciparum 3D7 MR4 |
| SA522686 | 4-12-AVQ_3 | Atovaquone | Plasmodium falciparum 3D7 MR4 |
| SA522687 | 4-8-Chloroquine_3 | Chloroquine | Plasmodium falciparum 3D7 MR4 |
| SA522688 | 4-8-Chloroquine_2 | Chloroquine | Plasmodium falciparum 3D7 MR4 |
| SA522689 | 4-8-Chloroquine_1 | Chloroquine | Plasmodium falciparum 3D7 MR4 |
| SA522690 | 4-12-Chloroquine_1 | Chloroquine | Plasmodium falciparum 3D7 MR4 |
| SA522691 | 4-12-Chloroquine_2 | Chloroquine | Plasmodium falciparum 3D7 MR4 |
| SA522692 | 4-12-Chloroquine_3 | Chloroquine | Plasmodium falciparum 3D7 MR4 |
| SA522693 | 4-13-Chloroquine_2 | Chloroquine | Plasmodium falciparum 3D7 MR4 |
| SA522694 | 4-13-Chloroquine_1 | Chloroquine | Plasmodium falciparum 3D7 MR4 |
| SA522695 | 4-13-Chloroquine_3 | Chloroquine | Plasmodium falciparum 3D7 MR4 |
| SA522696 | 4-12-DMSO_1 | DMSO Control | Plasmodium falciparum 3D7 MR4 |
| SA522697 | 4-12-DMSO_2 | DMSO Control | Plasmodium falciparum 3D7 MR4 |
| SA522698 | 4-13-DMSO_3 | DMSO Control | Plasmodium falciparum 3D7 MR4 |
| SA522699 | 4-13-DMSO_2 | DMSO Control | Plasmodium falciparum 3D7 MR4 |
| SA522700 | 4-13-DMSO_1 | DMSO Control | Plasmodium falciparum 3D7 MR4 |
| SA522701 | 4-8-DMSO_1 | DMSO Control | Plasmodium falciparum 3D7 MR4 |
| SA522702 | 4-8-DMSO_2 | DMSO Control | Plasmodium falciparum 3D7 MR4 |
| SA522703 | 4-8-DMSO_3 | DMSO Control | Plasmodium falciparum 3D7 MR4 |
| SA522704 | 4-12-DMSO_3 | DMSO Control | Plasmodium falciparum 3D7 MR4 |
| Showing results 1 to 45 of 45 |
Collection:
| Collection ID: | CO004578 |
| Collection Summary: | Each sample represents metabolites extracted from 1E8 Plasmodium falciparum infected human red blood cells (iRBCs). After treatment, the iRBCs were washed with cold PBS and metabolites were extracted using a 90% methanol solution. This solution was evaporated under nitrogen gas flow, resuspended in water plus chlorpropamide as an internal control, and run on the ThermoFisher Exactive Plus. |
| Sample Type: | Plasmodium cells |
Treatment:
| Treatment ID: | TR004594 |
| Treatment Summary: | Heparin synchronized (6-hour window) P. falciparum 3D7 parasites were grown to approximately 24 hours post-invasion, and infected red blood cells (iRBCs) were purified using magnet activated cell sorting (MACS). Purified iRBCs were then counted and plated at ~1X10^6 cells per technical replicate and allowed to recover in standard incubation conditions for 1.5 hours. After recovery, drug treatment was added at 10 X IC50 and incubated for 2.5 hours before washing and extraction of metabolites. |
Sample Preparation:
| Sampleprep ID: | SP004591 |
| Sampleprep Summary: | Parasites were pelleted (500 g, 4°C, 1 min) and washed in 1 mL ice cold PBS. Then metabolites were extracted from the pellet with 1 ml ice cold 90% methanol, vortexed 30 seconds, and centrifuged for 10 min at maximum speed (16000 x g) at 4°C. Samples were treated identically and swiftly to ensure reproducible results. The methanol supernatants were stored at -70°C until further processing could be done, when they were transferred to fresh 1.5 mL tubes, dried down completely under nitrogen gas flow, and the metabolite residues stored at -70°C. |
| Processing Storage Conditions: | On ice |
| Extraction Method: | 90% methanol |
| Extract Storage: | -80℃ |
Combined analysis:
| Analysis ID | AN007402 |
|---|---|
| Chromatography ID | CH005608 |
| MS ID | MS007094 |
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Thermo Dionex Ultimate 3000 |
| Column | Waters XSelect HSS T3 Column XP (100 x 2.1 mm, 2.5 µm) |
| MS Type | ESI |
| MS instrument type | Orbitrap |
| MS instrument name | Thermo Exactive Plus Orbitrap |
| Ion Mode | NEGATIVE |
| Units | blank-subtracted peak area |
Chromatography:
| Chromatography ID: | CH005608 |
| Instrument Name: | Thermo Dionex Ultimate 3000 |
| Column Name: | Waters XSelect HSS T3 Column XP (100 x 2.1 mm, 2.5 µm) |
| Column Temperature: | 30°C |
| Flow Gradient: | 0-5.0 min: 100% A, 0% B; 5.0-13.0 min: 80% A, 20% B; 13.0-15.0 min: 45% A, 55% B; 15.0-17.5 min: 35% A, 65% B; 17.5-21.0 min: 5% A, 95% B; 21.0-25 min: 100% A, 0% B |
| Flow Rate: | 0.200 mL/min |
| Internal Standard: | chlorpropamide 1 µM |
| Solvent A: | 97% water/3% methanol; 15 mM acetic acid; 10 mM tributylamine; 2.5 µM medronic acid |
| Solvent B: | 100% Methanol |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS007094 |
| Analysis ID: | AN007402 |
| Instrument Name: | Thermo Exactive Plus Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | .raw files were converted to .mzML files using MSConvert software of the ProteoWizard software package. The .mzML files were processed using EL MAVEN software for peak picking, alignment, and annotation. |
| Ion Mode: | NEGATIVE |
| Mass Accuracy: | 10 ppm |
| Resolution Setting: | 140,000 at m/z 200 |
| Scanning Range: | 85-1000 m/z |
