Molecular Target Validation of Aspartate Transcarbamoylase from Plasmodium falciparum by Torin 2

Autor(es)

Bosch, Soraya S. ^{[1, 2]} ; Lunev, Sergey ^[2] ; Batista, Fernando A. ^[2] ; Linzke, Marleen ^[1] ; Kronenberger, Thales ^[3] ; Domling, Alexander S. S. ^[2] ; Groves, Matthew R. ^[2] ; Wrenger, Carsten ^[1]

Afiliação do(s) autor(es)

^[1] Univ Sao Paulo, Inst Biomed Sci, Dept Parasitol, Unit Drug Discovery, BR-05508000 Sao Paulo, SP - Brazil

^[2] Univ Groningen, Dept Pharm, Struct Biol Unit, Drug Design XB20, NL-9700 AD Groningen - Netherlands

^[3] Univ Hosp Tubingen, Dept Internal Med 8, D-72076 Tubingen - Germany

• Tipo de documento: Artigo Científico
• Fonte: ACS INFECTIOUS DISEASES; v. 6, n. 5, p. 986-999, MAY 8 2020.
• Citações Web of Science: 0

Resumo

Malaria is a tropical disease that kills about half a million people around the world annually. Enzymatic reactions within pyrimidine biosynthesis have been proven to be essential for Plasmodium proliferation. Here we report on the essentiality of the second enzymatic step of the pyrimidine biosynthesis pathway, catalyzed by aspartate transcarbamoylase (ATC). Crystallization experiments using a double mutant of Plasmodium falciparum ATC (PfATC) revealed the importance of the mutated residues for enzyme catalysis. Subsequently, this mutant was employed in protein interference assays (PIAs), which resulted in inhibition of parasite proliferation when parasites transfected with the double mutant were cultivated in medium lacking an excess of nutrients, including aspartate. Addition of 5 or 10 mg/L of aspartate to the minimal medium restored the parasites' normal growth rate. In vitro and whole-cell assays in the presence of the compound Torin 2 showed inhibition of specific activity and parasite growth, respectively. In silico analyses revealed the potential binding mode of Torin 2 to PfATC. Furthermore, a transgenic ATC-overexpressing cell line exhibited a 10-fold increased tolerance to Torin 2 compared with control cultures. Taken together, our results confirm the antimalarial activity of Torin 2, suggesting PfATC as a target of this drug and a promising target for the development of novel antimalarials. (AU)

• Processo FAPESP: 17/03966-4 - Alvejando a via de recuperação e biossíntese de ácido lipoico em MRSA
• Beneficiário: Carsten Wrenger
• Linha de fomento: Auxílio à Pesquisa - Regular
• Processo FAPESP: 15/26722-8 - Drug discovery contra doenças infecciosas humanos
• Beneficiário: Carsten Wrenger
• Linha de fomento: Auxílio à Pesquisa - Temático
• Processo FAPESP: 13/17577-9 - Análise da catálise da ATCase no metabolismo de Plasmodium falciparum
• Beneficiário: Soraya Soledad Bosch
• Linha de fomento: Bolsas no Brasil - Doutorado Direto
• Processo FAPESP: 14/23330-9 - Análise morfológica da formação de apicoplasto em Plasmodium falciparum
• Beneficiário: Marleen Linzke
• Linha de fomento: Bolsas no Brasil - Doutorado