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

Author(s)

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]

Affiliation

^[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

• Document type: Journal article
• Source: ACS INFECTIOUS DISEASES; v. 6, n. 5, p. 986-999, MAY 8 2020.
• Web of Science Citations: 0

Abstract

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)

• FAPESP's process: 17/03966-4 - Targeting lipoic acid salvage and biosynthesis pathways in MRSA
• Grantee: Carsten Wrenger
• Support Opportunities: Regular Research Grants
• FAPESP's process: 15/26722-8 - Drug discovery against human infectious diseases
• Grantee: Carsten Wrenger
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 13/17577-9 - Analysis of the ATCase catalysis within the amino acid metabolism of the human malaria parasite Plasmodium falciparum
• Grantee: Soraya Soledad Bosch
• Support Opportunities: Scholarships in Brazil - Doctorate (Direct)
• FAPESP's process: 14/23330-9 - Morphologic Analysis of the Apicoplast Formation in Plasmodium falciparum
• Grantee: Marleen Linzke
• Support Opportunities: Scholarships in Brazil - Doctorate