Proteomics

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Molecular markers of adaptive responses in Toxoplasma gondii to Pyrimethamine treatment

ABSTRACT: In this study, we employed bio-orthogonal non-canonical amino acid tagging (BONCAT) combined with iTRAQ-based quantitative proteomics to investigate the effects of pyrimethamine (PM) on newly synthesised proteins (NSPs) in Toxoplasma gondii. Parasites were treated with varying concentrations of PM, followed by mass spectrometric analysis to identify and quantify NSPs. We observed a concentration-dependent response, where several NSPs were downregulated at low PM doses but upregulated at higher concentrations. Notably, ribosomal proteins, invasion-related proteins (e.g., ROP26), and stress response proteins (e.g., serine/threonine-protein phosphatase 2B) showed significant changes in expression. The upregulation of calcineurin-related proteins suggests an activation of calcium-dependent stress response pathways, possibly as a compensatory mechanism to counteract the drug’s inhibitory effects. Our findings reveal the dynamic regulatory mechanisms of T. gondii under PM treatment, highlighting the parasite’s adaptive strategies to survive drug-induced stress. This study underscores the utility of BONCAT-iTRAQ proteomics in uncovering the molecular responses of pathogens to therapeutic interventions, providing insights that could inform the development of more effective anti-parasitic strategies.

INSTRUMENT(S): TripleTOF 5600

ORGANISM(S): Toxoplasma Gondii

SUBMITTER: Karunakaran Kalesh  

LAB HEAD: Kalesh Karunakaran

PROVIDER: PXD054994 | Pride | 2025-05-07

REPOSITORIES: Pride

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BONCAT-iTRAQ Labelling Reveals Molecular Markers of Adaptive Responses in <i>Toxoplasma gondii</i> to Pyrimethamine Treatment.

Mina John G JG   Parthasarathy Anutthaman A   Porta Exequiel O EO   Denny Paul W PW   Kalesh Karunakaran K  

Pathogens (Basel, Switzerland) 20241008 10

We employed a BONCAT-iTRAQ labelling approach to investigate newly synthesised proteins (NSPs) in <i>Toxoplasma gondii</i> subjected to varying concentrations of the antifolate drug pyrimethamine. Our results reveal that numerous NSPs exhibited altered expression levels in response to the drug, with significant upregulation observed at higher concentrations. Key proteins involved in protein synthesis, stress responses, energy metabolism, and cytoskeletal dynamics were identified, indicating that  ...[more]

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