System-wide biochemical analysis reveals ozonide antimalarials initially act by disrupting Plasmodium falciparum haemoglobin digestion.
Ozonide antimalarials, OZ277 (arterolane) and OZ439 (artefenomel), are synthetic peroxide-based antimalarials with potent activity against the deadliest malaria parasite, Plasmodium falciparum. Here we used a "multi-omics" workflow, in combination with activity-based protein profiling (ABP...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2020-06-01
|
| Series: | PLoS Pathogens |
| Online Access: | https://doi.org/10.1371/journal.ppat.1008485 |
| _version_ | 1819260860092121088 |
|---|---|
| author | Carlo Giannangelo Ghizal Siddiqui Amanda De Paoli Bethany M Anderson Laura E Edgington-Mitchell Susan A Charman Darren J Creek |
| author_facet | Carlo Giannangelo Ghizal Siddiqui Amanda De Paoli Bethany M Anderson Laura E Edgington-Mitchell Susan A Charman Darren J Creek |
| author_sort | Carlo Giannangelo |
| collection | DOAJ |
| description | Ozonide antimalarials, OZ277 (arterolane) and OZ439 (artefenomel), are synthetic peroxide-based antimalarials with potent activity against the deadliest malaria parasite, Plasmodium falciparum. Here we used a "multi-omics" workflow, in combination with activity-based protein profiling (ABPP), to demonstrate that peroxide antimalarials initially target the haemoglobin (Hb) digestion pathway to kill malaria parasites. Time-dependent metabolomic profiling of ozonide-treated P. falciparum infected red blood cells revealed a rapid depletion of short Hb-derived peptides followed by subsequent alterations in lipid and nucleotide metabolism, while untargeted peptidomics showed accumulation of longer Hb-derived peptides. Quantitative proteomics and ABPP assays demonstrated that Hb-digesting proteases were increased in abundance and activity following treatment, respectively. Ozonide-induced depletion of short Hb-derived peptides was less extensive in a drug-treated K13-mutant artemisinin resistant parasite line (Cam3.IIR539T) than in the drug-treated isogenic sensitive strain (Cam3.IIrev), further confirming the association between ozonide activity and Hb catabolism. To demonstrate that compromised Hb catabolism may be a primary mechanism involved in ozonide antimalarial activity, we showed that parasites forced to rely solely on Hb digestion for amino acids became hypersensitive to short ozonide exposures. Quantitative proteomics analysis also revealed parasite proteins involved in translation and the ubiquitin-proteasome system were enriched following drug treatment, suggestive of the parasite engaging a stress response to mitigate ozonide-induced damage. Taken together, these data point to a mechanism of action involving initial impairment of Hb catabolism, and indicate that the parasite regulates protein turnover to manage ozonide-induced damage. |
| first_indexed | 2024-12-23T19:32:37Z |
| format | Article |
| id | doaj.art-82eb7953137945c3bc3295e0874b763d |
| institution | Directory Open Access Journal |
| issn | 1553-7366 1553-7374 |
| language | English |
| last_indexed | 2024-12-23T19:32:37Z |
| publishDate | 2020-06-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Pathogens |
| spelling | doaj.art-82eb7953137945c3bc3295e0874b763d2022-12-21T17:33:52ZengPublic Library of Science (PLoS)PLoS Pathogens1553-73661553-73742020-06-01166e100848510.1371/journal.ppat.1008485System-wide biochemical analysis reveals ozonide antimalarials initially act by disrupting Plasmodium falciparum haemoglobin digestion.Carlo GiannangeloGhizal SiddiquiAmanda De PaoliBethany M AndersonLaura E Edgington-MitchellSusan A CharmanDarren J CreekOzonide antimalarials, OZ277 (arterolane) and OZ439 (artefenomel), are synthetic peroxide-based antimalarials with potent activity against the deadliest malaria parasite, Plasmodium falciparum. Here we used a "multi-omics" workflow, in combination with activity-based protein profiling (ABPP), to demonstrate that peroxide antimalarials initially target the haemoglobin (Hb) digestion pathway to kill malaria parasites. Time-dependent metabolomic profiling of ozonide-treated P. falciparum infected red blood cells revealed a rapid depletion of short Hb-derived peptides followed by subsequent alterations in lipid and nucleotide metabolism, while untargeted peptidomics showed accumulation of longer Hb-derived peptides. Quantitative proteomics and ABPP assays demonstrated that Hb-digesting proteases were increased in abundance and activity following treatment, respectively. Ozonide-induced depletion of short Hb-derived peptides was less extensive in a drug-treated K13-mutant artemisinin resistant parasite line (Cam3.IIR539T) than in the drug-treated isogenic sensitive strain (Cam3.IIrev), further confirming the association between ozonide activity and Hb catabolism. To demonstrate that compromised Hb catabolism may be a primary mechanism involved in ozonide antimalarial activity, we showed that parasites forced to rely solely on Hb digestion for amino acids became hypersensitive to short ozonide exposures. Quantitative proteomics analysis also revealed parasite proteins involved in translation and the ubiquitin-proteasome system were enriched following drug treatment, suggestive of the parasite engaging a stress response to mitigate ozonide-induced damage. Taken together, these data point to a mechanism of action involving initial impairment of Hb catabolism, and indicate that the parasite regulates protein turnover to manage ozonide-induced damage.https://doi.org/10.1371/journal.ppat.1008485 |
| spellingShingle | Carlo Giannangelo Ghizal Siddiqui Amanda De Paoli Bethany M Anderson Laura E Edgington-Mitchell Susan A Charman Darren J Creek System-wide biochemical analysis reveals ozonide antimalarials initially act by disrupting Plasmodium falciparum haemoglobin digestion. PLoS Pathogens |
| title | System-wide biochemical analysis reveals ozonide antimalarials initially act by disrupting Plasmodium falciparum haemoglobin digestion. |
| title_full | System-wide biochemical analysis reveals ozonide antimalarials initially act by disrupting Plasmodium falciparum haemoglobin digestion. |
| title_fullStr | System-wide biochemical analysis reveals ozonide antimalarials initially act by disrupting Plasmodium falciparum haemoglobin digestion. |
| title_full_unstemmed | System-wide biochemical analysis reveals ozonide antimalarials initially act by disrupting Plasmodium falciparum haemoglobin digestion. |
| title_short | System-wide biochemical analysis reveals ozonide antimalarials initially act by disrupting Plasmodium falciparum haemoglobin digestion. |
| title_sort | system wide biochemical analysis reveals ozonide antimalarials initially act by disrupting plasmodium falciparum haemoglobin digestion |
| url | https://doi.org/10.1371/journal.ppat.1008485 |
| work_keys_str_mv | AT carlogiannangelo systemwidebiochemicalanalysisrevealsozonideantimalarialsinitiallyactbydisruptingplasmodiumfalciparumhaemoglobindigestion AT ghizalsiddiqui systemwidebiochemicalanalysisrevealsozonideantimalarialsinitiallyactbydisruptingplasmodiumfalciparumhaemoglobindigestion AT amandadepaoli systemwidebiochemicalanalysisrevealsozonideantimalarialsinitiallyactbydisruptingplasmodiumfalciparumhaemoglobindigestion AT bethanymanderson systemwidebiochemicalanalysisrevealsozonideantimalarialsinitiallyactbydisruptingplasmodiumfalciparumhaemoglobindigestion AT lauraeedgingtonmitchell systemwidebiochemicalanalysisrevealsozonideantimalarialsinitiallyactbydisruptingplasmodiumfalciparumhaemoglobindigestion AT susanacharman systemwidebiochemicalanalysisrevealsozonideantimalarialsinitiallyactbydisruptingplasmodiumfalciparumhaemoglobindigestion AT darrenjcreek systemwidebiochemicalanalysisrevealsozonideantimalarialsinitiallyactbydisruptingplasmodiumfalciparumhaemoglobindigestion |