| Texto completo | |
| Autor(es): Mostrar menos - |
Marcelino, Tiago de Paula
;
Fala, Angela Maria
;
Silva, Matheus Monteiro da
;
Souza-Melo, Normanda
;
Malvezzi, Amaranta Muniz
;
Klippel, Angelica Hollunder
;
Zoltner, Martin
;
Padilla-Mejia, Norma
;
Kosto, Samantha
;
Field, Mark C.
;
Burle-Caldas, Gabriela de Assis
;
Teixeira, Santuza Maria Ribeiro
;
Counago, Rafael Miguez
;
Massirer, Katlin Brauer
;
Schenkman, Sergio
Número total de Autores: 15
|
| Tipo de documento: | Artigo Científico |
| Fonte: | Journal of Biological Chemistry; v. 299, n. 7, p. 18-pg., 2023-07-01. |
| Resumo | |
The TcK2 protein kinase of Trypanosoma cruzi, the causative agent of Chagas disease, is structurally similar to the human kinase PERK, which phosphorylates the initiation factor eIF2 alpha and, in turn, inhibits translation initiation. We have previously shown that absence of TcK2 kinase impairs parasite proliferation within mammalian cells, positioning it as a potential target for treatment of Chagas disease. To better understand its role in the parasite, here we initially confirmed the importance of TcK2 in parasite proliferation by generating CRISPR/Cas9 TcK2-null cells, albeit they more efficiently differentiate into infective forms. Proteomics indicates that the TcK2 knockout of proliferative forms expresses proteins including trans-sialidases, normally restricted to infective and nonproliferative trypomastigotes explaining decreased proliferation and better differentiation. TcK2 knockout cells lost phosphorylation of eukaryotic initiation factor 3 and cyclic AMP responsive-like element, recognized to promote growth, likely explaining both decreased proliferation and augmented differentiation. To identify specific inhibitors, a library of 379 kinase inhibitors was screened by differential scanning fluo- rimetry using a recombinant TcK2 encompassing the kinase domain and selected molecules were tested for kinase inhibition. Only Dasatinib and PF -477736, inhibitors of Src/Abl and ChK1 kinases, showed inhibitory activity with IC50 of 0.2 +/- 0.02 mM and 0.8 +/- 0.1, respectively. In infected cells Dasatinib inhibited growth of parental amastigotes (IC50 = 0.6 +/- 0.2 mM) but not TcK2 of depleted parasites (IC50 > 34 mM) identifying Dasatinib as a potential lead for development of therapeutics for Chagas disease targeting TcK2. (AU) | |
| Processo FAPESP: | 17/02496-4 - Papel da acetilação de proteínas e sirtuínas no processo de infecção pelo T. cruzi |
| Beneficiário: | Amaranta Muniz Malvezzi |
| Modalidade de apoio: | Bolsas no Brasil - Pós-Doutorado |
| Processo FAPESP: | 20/07870-4 - Mecanismos de adaptação de tripanossomatídeos ao hospedeiro através de controle da transcrição, síntese proteica e secreção de vesículas extracelulares |
| Beneficiário: | Ana Claudia Trocoli Torrecilhas |
| Modalidade de apoio: | Auxílio à Pesquisa - Temático |
| Processo FAPESP: | 19/15909-0 - Estudos do mecanismos de secreção de vesículas extracelulares por protozoários parasitas |
| Beneficiário: | Sergio Schenkman |
| Modalidade de apoio: | Auxílio à Pesquisa - Regular |
| Processo FAPESP: | 14/50897-0 - INCT 2014: Centro de Química Medicinal de Acesso Aberto |
| Beneficiário: | Katlin Brauer Massirer |
| Modalidade de apoio: | Auxílio à Pesquisa - Temático |
| Processo FAPESP: | 20/02006-0 - Caracterização molecular e estrutural de proteínas de ligação a RNA como alvos terapêuticos em câncer e neurologia |
| Beneficiário: | Katlin Brauer Massirer |
| Modalidade de apoio: | Auxílio à Pesquisa - Parceria para Inovação Tecnológica - PITE |
| Processo FAPESP: | 17/02416-0 - Secreção de proteínas em Trypanosoma |
| Beneficiário: | Normanda Souza Melo |
| Modalidade de apoio: | Bolsas no Brasil - Pós-Doutorado |