Summary of Study ST004290
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 PR002709. The data can be accessed directly via it's Project DOI: 10.21228/M8GV8W This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.
| Study ID | ST004290 |
| Study Title | Metabolomics characterisation of Plasmodium falciparum response to plasmepsin V peptidomimetic inhibitors - 5 hour treatment |
| Study Summary | 3D7 Plasmodium falciparum parasites were treated with plasmepsin V peptidometics (5h) to check for pertrubation of metabolism. Following treatment, parasites were harvested and metabolism analysed. It was observed that following 5 h incubation, parasites changed their peptide signature and was concluded that these inhibitors may perturb peptide metabolism. |
| Institute | Monash University |
| Last Name | Siddiqui |
| First Name | Ghizal |
| Address | 381 Royal Parade, Parkville, Melbourne, Victoria, 3052, Australia |
| ghizal.siddiqui@monash.edu | |
| Phone | 99039282 |
| Submit Date | 2025-10-12 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | mzML, raw(Thermo) |
| Analysis Type Detail | LC-MS |
| Release Date | 2025-11-03 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR002709 |
| Project DOI: | doi: 10.21228/M8GV8W |
| Project Title: | Deconvolution of the on-target activity of plasmepsin V peptidomimetics in P. falciparum parasites |
| Project Summary: | Plasmepsin V (PMV), an essential aspartyl protease, plays a critical role during the asexual blood stage of infection of Plasmodium by enabling the export of parasite proteins into the host red blood cell. This export is vital for parasite survival and pathogenesis, making PMV an attractive target for antimalarial drug development. Peptidomimetic inhibitors designed to mimic the natural substrate of PMV have demonstrated potent parasite-killing activity by blocking protein export. While these compounds have been instrumental in validating PMV as a bona fide antimalarial target, inconsistencies between their biochemical potency and cellular activity have raised questions regarding their precise mechanism of action. In this study, we employed chemoproteomic approaches, including solvent induced protein precipitation and intact-cell thermal profiling, to demonstrate PMV target engagement by the peptidomimetics. To further support these findings, we generated parasite lines exhibiting reduced sensitivity to peptidomimetics. Through whole-genome sequencing of these parasite lines, a single nucleotide variant within the pmv gene was revealed. This mutation was later validated using reverse genetics, confirming its role in mediating resistance. Together, these data provide strong evidence that the peptidomimetics exert their antimalarial activity by directly targeting PMV. These findings further support the potential of PMV as a validated and promising target for future antimalarial drug development. |
| Institute: | Monash University |
| Last Name: | Siddiqui |
| First Name: | Ghizal |
| Address: | 381 Royal Parade, Parkville, Melbourne, Victoria, 3052, Australia |
| Email: | ghizal.siddiqui@monash.edu |
| Phone: | 99039282 |
Subject:
| Subject ID: | SU004443 |
| Subject Type: | Cultured cells |
| Subject Species: | Plasmodium falciparum |
| Taxonomy ID: | 5833 |
Factors:
Subject type: Cultured cells; Subject species: Plasmodium falciparum (Factor headings shown in green)
| mb_sample_id | local_sample_id | Sample source | factor |
|---|---|---|---|
| SA498567 | P_E23_I_DMSO_3_5_h | iRBC | DMSO |
| SA498568 | P_E23_I_DMSO_3_4_g | iRBC | DMSO |
| SA498569 | P_E23_I_DMSO_3_3_f | iRBC | DMSO |
| SA498570 | P_E23_I_DMSO_3_2_e | iRBC | DMSO |
| SA498571 | P_E23_I_DMSO_3_1_d | iRBC | DMSO |
| SA498572 | P_E23_I_DMSO_2_c | iRBC | DMSO |
| SA498573 | P_E23_I_DMSO_1_2_b | iRBC | DMSO |
| SA498574 | P_E23_I_DMSO_1_1_a | iRBC | DMSO |
| SA498575 | P_E23_I_024_1_2_b | iRBC | WEHI-024 |
| SA498576 | P_E23_I_024_1_1_a | iRBC | WEHI-024 |
| SA498577 | P_E23_I_024_3_4_g | iRBC | WEHI-024 |
| SA498578 | P_E23_I_024_3_5_h | iRBC | WEHI-024 |
| SA498579 | P_E23_I_024_3_3_f | iRBC | WEHI-024 |
| SA498580 | P_E23_I_024_3_2_e | iRBC | WEHI-024 |
| SA498581 | P_E23_I_024_3_1_d | iRBC | WEHI-024 |
| SA498582 | P_E23_I_024_2_c | iRBC | WEHI-024 |
| SA498583 | P_E23_I_601_3_1_d | iRBC | WEHI-601 |
| SA498584 | P_E23_I_601_2_c | iRBC | WEHI-601 |
| SA498585 | P_E23_I_601_3_3_f | iRBC | WEHI-601 |
| SA498586 | P_E23_I_601_3_4_g | iRBC | WEHI-601 |
| SA498587 | P_E23_I_601_3_5_h | iRBC | WEHI-601 |
| SA498588 | P_E23_I_601_1_2_b | iRBC | WEHI-601 |
| SA498589 | P_E23_I_601_1_1_a | iRBC | WEHI-601 |
| SA498590 | P_E23_I_601_3_2_e | iRBC | WEHI-601 |
| Showing results 1 to 24 of 24 |
Collection:
| Collection ID: | CO004436 |
| Collection Summary: | Pf3D7 Plasmodium falciparum infected red blood cells (iRBCs) were grown in RPMI medium containing hypoxanthine and 0.5% (wt/vol) Albumax (Gibco). For the 5-h treatment, parasites were synchronised by double 5% (wt/vol) sorbitol lysis, 14 h apart and cultured for a further 52 h to ensure the experiment was performed on asexual early-trophozoite stage cultures ∼22-24 hpi. |
| Sample Type: | Blood (whole) |
Treatment:
| Treatment ID: | TR004452 |
| Treatment Summary: | Parasites were adjusted to 6% parasitaemia and treated with (Walter and Eliza Hall Institute) WEHI-601 or WEHI-024 (0.9 µM) for 5 h. Following the treatment duration, parasites were harvested using magnet separation to achieve enriched trophozoite samples (>80% parasitaemia). Untreated controls contained equivalent amounts of dimethyl sulfoxide (DMSO; as a vehicle). |
Sample Preparation:
| Sampleprep ID: | SP004449 |
| Sampleprep Summary: | After drug treatments, metabolites of infected RBCs (iRBCs) were collected as previously described (doi: 10.1128/AAC.01226-16), with minor modifications. Metabolites of iRBCs in the 5 h treatments were extracted with of methanol, respectively. |
Chromatography:
| Chromatography ID: | CH005420 |
| Chromatography Summary: | Metabolite analysis was performed by liquid chromatography-mass spectrometry LC-MS using hydrophilic interaction liquid chromatography (HILIC) and high-resolution (Q-Exactive Orbitrap, Thermo Fisher) MS. Briefly, samples (10 μL) were injected onto a Dionex RSLC U3000 LC system (Thermo) fitted with a ZIC-pHILIC column (5 μm particle size, 4.6 by 150 mm; Merck) and 20 mM ammonium carbonate (A) and acetonitrile (B) were used as the mobile phases. A 30 min gradient starting from 80% B to 50% B over 15 mins, followed by washing at 5% B for 3 mins and re-equilibration at 80% B, was used. |
| Instrument Name: | Thermo Dionex Ultimate 3000 RS |
| Column Name: | Merck SeQuant ZIC-pHILIC (150 x 4.6mm,5um) |
| Column Temperature: | 25 |
| Flow Gradient: | A 30 min gradient starting from 80% B to 50% B over 15 mins, followed by washing at 5% B for 3 mins and re-equilibration at 80% B, was used. |
| Flow Rate: | 0.35 ml/min |
| Solvent A: | 100% water; 20 mM ammonium carbonate |
| Solvent B: | 100% Acetonitrile |
| Chromatography Type: | HILIC |
Analysis:
| Analysis ID: | AN007133 |
| Analysis Type: | MS |
| Chromatography ID: | CH005420 |
| Num Factors: | 3 |
| Num Metabolites: | 157 |
| Units: | normalised ion counts |
| Analysis ID: | AN007134 |
| Analysis Type: | MS |
| Chromatography ID: | CH005420 |
| Num Factors: | 3 |
| Num Metabolites: | 215 |
| Units: | normalised ion counts |
