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

Autor(es)

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]}

Afiliação do(s) autor(es)

^[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

• Tipo de documento: Artigo Científico
• Fonte: ACS Chemical Biology; v. 15, n. 7, p. 1801-1807, JUL 17 2020.
• Citações Web of Science: 0

Resumo

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)

• Processo FAPESP: 13/50724-5 - Centro de Biologia Química de Proteínas Quinases: alavancando desenvolvimento de fármacos através de pesquisa de acesso aberto
• Beneficiário: Paulo Arruda
• Modalidade de apoio: Auxílio à Pesquisa - Parceria para Inovação Tecnológica - PITE
• Processo FAPESP: 16/17469-0 - Desenvolvimento de ensaios celulares para avaliar e quantificar as alterações fenotípicas da inibição química da Vaccinia-Related Kinase 1 (VRK1)
• Beneficiário: Roberta Regina Ruela de Sousa
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado