Targeting prolyl-tRNA synthetase via a series of ATP-mimetics to accelerate drug discovery against toxoplasmosis

DOI: 10.1371/journal.ppat.1011124

Authors: Manickam Yogavel (International Centre for Genetic Engineering and Biotechnology (ICGEB)) , Alexandre Bougdour (INSERM U1209, CNRS UMR5309, Université Grenoble Alpes) , Siddhartha Mishra (nternational Centre for Genetic Engineering and Biotechnology (ICGEB); Academy of Scientific and Innovative Research (AcSIR); ICMR-National Institute of Malaria Research) , Nipun Malhotra (International Centre for Genetic Engineering and Biotechnology (ICGEB)) , Jyoti Chhibber-Goel (International Centre for Genetic Engineering and Biotechnology (ICGEB)) , Valeria Bellini (NSERM U1209, CNRS UMR5309, Université Grenoble Alpes) , Karl Harlos (Wellcome Trust Centre for Human Genetics, University of Oxford) , Benoît Laleu (Medicines for Malaria Venture (MMV), International Center Cointrin (ICC)) , Mohamed-Ali Hakimi (INSERM U1209, CNRS UMR5309, Université Grenoble Alpes) , Amit Sharma (International Centre for Genetic Engineering and Biotechnology (ICGEB); Academy of Scientific and Innovative Research (AcSIR); ICMR-National Institute of Malaria Research)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Plos Pathogens , VOL 19

State: Published (Approved)
Published: February 2023
Diamond Proposal Number(s): 14744

Abstract: The prolyl-tRNA synthetase (PRS) is a validated drug target for febrifugine and its synthetic analog halofuginone (HFG) against multiple apicomplexan parasites including Plasmodium falciparum and Toxoplasma gondii. Here, a novel ATP-mimetic centered on 1-(pyridin-4-yl) pyrrolidin-2-one (PPL) scaffold has been validated to bind to Toxoplasma gondii PRS and kill toxoplasma parasites. PPL series exhibited potent inhibition at the cellular (T. gondii parasites) and enzymatic (TgPRS) levels compared to the human counterparts. Cell-based chemical mutagenesis was employed to determine the mechanism of action via a forward genetic screen. Tg-resistant parasites were analyzed with wild-type strain by RNA-seq to identify mutations in the coding sequence conferring drug resistance by computational analysis of variants. DNA sequencing established two mutations, T477A and T592S, proximal to terminals of the PPL scaffold and not directly in the ATP, tRNA, or L-pro sites, as supported by the structural data from high-resolution crystal structures of drug-bound enzyme complexes. These data provide an avenue for structure-based activity enhancement of this chemical series as anti-infectives.

Journal Keywords: Toxoplasma gondii; Parasitic diseases; Enzyme structure; Crystal structure; RNA sequencing; Enzyme inhibitors; Insertion mutation; Drug interactions

Diamond Keywords: Toxoplasmosis; Enzymes

Subject Areas: Biology and Bio-materials, Medicine


Instruments: I04-Macromolecular Crystallography , I24-Microfocus Macromolecular Crystallography

Other Facilities: PROIXMA 1, PROXIMA 2A at SOLEIL

Added On: 06/03/2023 09:25

Documents:
journal.ppat.1011124.pdf

Discipline Tags:

Infectious Diseases Health & Wellbeing Structural biology Drug Discovery Life Sciences & Biotech Parasitology

Technical Tags:

Diffraction Macromolecular Crystallography (MX)