Proteomics

Dataset Information

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Thermal proteome profiling of Toxoplasma gondii lysates treated with enhancer 1 (ENH1)


ABSTRACT: Previously, we characterized a chemical modulator, ENH1, that exhibited potent antiparasitic properties against apicomplexans and induced calcium oscillations, but the target eluded identification. To determine the target of ENH1, we employed thermal proteome profiling in parasites. We treated extracellular parasites with vehicle (DMSO) or ENH1 for 15 minutes at 37°C, under conditions previously determined to induce calcium fluxes in T. gondii.8 Parasites were then heated at 10 different temperatures to induce thermal denaturation. Following lysis and high-speed centrifugation, soluble protein was isolated and analyzed by mass spectrometry to generate thermal stability profiles from each condition across two biological replicates.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Toxoplasma Gondii Gt1

SUBMITTER: Alice Herneisen  

LAB HEAD: Sebastian Lourido

PROVIDER: PXD018810 | Pride | 2020-07-13

REPOSITORIES: Pride

Dataset's files

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Publications

Identifying the Target of an Antiparasitic Compound in <i>Toxoplasma</i> Using Thermal Proteome Profiling.

Herneisen Alice L AL   Sidik Saima M SM   Markus Benedikt M BM   Drewry David H DH   Zuercher William J WJ   Lourido Sebastian S  

ACS chemical biology 20200706 7


Apicomplexan parasites include the causative agents of malaria and toxoplasmosis. Cell-based screens in <i>Toxoplasma</i> previously identified a chemical modulator of calcium signaling (ENH1) that blocked parasite egress from host cells and exhibited potent antiparasitic activity. To identify the targets of ENH1, we adapted thermal proteome profiling to <i>Toxoplasma</i>, which revealed calcium-dependent protein kinase 1 (CDPK1) as a target. We confirmed the inhibition of CDPK1 by ENH1 in vitro  ...[more]

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