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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Identifying the Target of an Antiparasitic Compound in Toxoplasma Using Thermal Proteome Profiling

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Herneisen, Alice L. [1, 2] ; Sidik, Saima M. [1] ; Markus, Benedikt M. [3, 1] ; Drewry, David H. [4, 5] ; Zuercher, William J. [4] ; Lourido, Sebastian [1, 2]
Total Authors: 6
[1] Whitehead Inst Biomed Res, 9 Cambridge Ctr, Cambridge, MA 02142 - USA
[2] MIT, Dept Biol, 77 Massachusetts Ave, Cambridge, MA 02142 - USA
[3] Univ Freiburg, Fac Biol, D-79104 Freiburg - Germany
[4] Univ N Carolina, Struct Genom Consortium, UNC Eshelman Sch Pharm, Chapel Hill, NC 27599 - USA
[5] Univ N Carolina, Div Chem Biol & Med Chem, UNC Eshelman Sch Pharm, Chapel Hill, NC 27599 - USA
Total Affiliations: 5
Document type: Journal article
Source: ACS Chemical Biology; v. 15, n. 7, p. 1801-1807, JUL 17 2020.
Web of Science Citations: 0

Apicomplexan parasites include the causative agents of malaria and toxoplasmosis. Cell-based screens in Toxoplasma 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 Toxoplasma, which revealed calcium-dependent protein kinase 1 (CDPK1) as a target. We confirmed the inhibition of CDPK1 by ENH1 in vitro and in parasites by comparing alleles sensitive or resistant to ENH1. CDPK1 inhibition explained the block in egress; however, the effects of ENH1 on calcium homeostasis and parasite viability were CDPK1-independent, implicating additional targets. Thermal proteome profiling of lysates from parasites expressing the resistant allele of CDPK1 identified additional candidates associated with the mitochondria and the parasite pellicle-compartments that potentially function in calcium release and homeostasis. Our findings illustrate the promise of thermal profiling to identify druggable targets that modulate calcium signaling in apicomplexan parasites. (AU)

FAPESP's process: 13/50724-5 - Protein Kinase Chemical Biology Center: supporting drug development through open-access research
Grantee:Paulo Arruda
Support Opportunities: Research Grants - Research Partnership for Technological Innovation - PITE
FAPESP's process: 16/17469-0 - Development of celular assays to measure and to discover the phenotypic consequences of chemical inhibition of Vaccinia-Related Kinase 1 (VRK1)
Grantee:Roberta Regina Ruela de Sousa
Support Opportunities: Scholarships in Brazil - Post-Doctoral