6HL7

Crystal structure of truncated aspartate transcarbamoylase from Plasmodium falciparum with mutated active site (R109A/K138A) and N-carbamoyl-L-phosphate bound

Summary for 6HL7

• Entry DOI: 10.2210/pdb6hl7/pdb
• Descriptor: Aspartate transcarbamoylase, PHOSPHORIC ACID MONO(FORMAMIDE)ESTER (3 entities in total)
• Functional Keywords: falciparum malaria pyrimidine biosynthesis trimer mutant transferase carbamoyl-phospahte, transferase
• Biological source: Plasmodium falciparum (malaria parasite P. falciparum)
• Total number of polymer chains: 3
• Total formula weight: 120640.43

Authors

Bosch, S.S.,Lunev, S.,Wrenger, C.,Groves, M.R. (deposition date: 2018-09-10, release date: 2018-09-26, Last modification date: 2024-01-24)

Primary citation

Bosch, S.S.,Lunev, S.,Batista, F.A.,Linzke, M.,Kronenberger, T.,Domling, A.S.S.,Groves, M.R.,Wrenger, C.
Molecular Target Validation of Aspartate Transcarbamoylase fromPlasmodium falciparumby Torin 2.
Acs Infect Dis., 6:986-999, 2020

PubMed 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 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 ATC (ATC) 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. and whole-cell assays in the presence of the compound Torin 2 showed inhibition of specific activity and parasite growth, respectively. analyses revealed the potential binding mode of Torin 2 to ATC. 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 ATC as a target of this drug and a promising target for the development of novel antimalarials.

PubMed: 32129597
DOI: 10.1021/acsinfecdis.9b00411

Experimental method

X-RAY DIFFRACTION (2.5 Å)