Rapid Chagas Disease Drug Target Discovery Using Directed Evolution in Drug-Sensitive Yeast

University of California. School of Medicine. Department of Pediatrics. San Diego, CA, USA.
University of California. School of Medicine. Department of Pediatrics. San Diego, CA, USA / J. Craig Venter Institute. Department of Synthetic Biology and Bioenergy. La Jolla, CA, USA.
University of California San Diego. Skaggs School of Pharmacy and Pharmaceutical Sciences. La Jolla, CA, USA / Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Ultraestrutura Celular. Rio de Janeiro, RJ, Brasil
University of California San Diego. Skaggs School of Pharmacy and Pharmaceutical Sciences. La Jolla, CA, USA.
University of California. School of Medicine. Department of Pediatrics. San Diego, CA, USA.
University of California. School of Medicine. Department of Pediatrics. San Diego, CA, USA.
University of California. School of Medicine. Department of Pediatrics. San Diego, CA, USA.
University of California. School of Medicine. Department of Pediatrics. San Diego, CA, USA.
University of California San Diego. Skaggs School of Pharmacy and Pharmaceutical Sciences. La Jolla, CA, USA.
University of California. School of Medicine. Department of Pediatrics. San Diego, CA, USA / Universidade Federal do Paraná. Departamento de Farmácia. Curitiba, PR, Brasil
University of California. School of Medicine. Department of Pediatrics. San Diego, CA, USA.
University of California. School of Medicine. Department of Pediatrics. San Diego, CA, USA.
J. Craig Venter Institute. Department of Synthetic Biology and Bioenergy. La JOlla, CA, USA.
University of California San Diego. Skaggs School of Pharmacy and Pharmaceutical Sciences. La Jolla, CA, USA.
University of California San Diego. Skaggs School of Pharmacy and Pharmaceutical Sciences. La Jolla, CA, USA.
University of California. Department of Chemistry & Biochemistry. San Diego, La Jolla, CA, USA.
University of California San Diego. Skaggs School of Pharmacy and Pharmaceutical Sciences. La Jolla, CA, USA.
University of California. Department of Chemistry & Biochemistry. San Diego, La Jolla, CA, USA / University of Pittsburgh. Department of Biological Sciences. Piittsburg, Pensylvania, USA.
University of California. School of Medicine. Department of Pediatrics. San Diego, CA, USA.

Abstract

Recent advances in cell-based, high-throughput phenotypic screening have identified new chemical compounds that are active against eukaryotic pathogens. A challenge to their future development lies in identifying these compounds' molecular targets and binding modes. In particular, subsequent structure-based chemical optimization and target-based screening require a detailed understanding of the binding event. Here, we use directed evolution and whole-genome sequencing of a drug-sensitive S. cerevisiae strain to identify the yeast ortholog of TcCyp51, lanosterol-14-alpha-demethylase (TcCyp51), as the target of MMV001239, a benzamide compound with activity against Trypanosoma cruzi, the etiological agent of Chagas disease. We show that parasites treated with MMV0001239 phenocopy parasites treated with another TcCyp51 inhibitor, posaconazole, accumulating both lanosterol and eburicol. Direct drug-protein binding of MMV0001239 was confirmed through spectrophotometric binding assays and X-ray crystallography, revealing a binding site shared with other antitrypanosomal compounds that target Cyp51. These studies provide a new probe chemotype for TcCyp51 inhibition.

Publisher

American Chemical Society

Citation

OTTILIE, Sabine; et al. Rapid Chagas disease drug target discovery using directed evolution in drug-sensitive yeast. ACS Chem Biol., v.12, n.2, p.422–434, Feb. 2017.

DOI

10.1021/acschembio.6b01037

ISSN

1554-8929

Please use this identifier to cite or link to this item: https://www.arca.fiocruz.br/handle/icict/23183

Type

Copyright

Restricted access

Embargo date

2030-01-01

Sustainable Development Goals

Collections

Share

Statistics

Metadata

Author

Affilliation

Abstract

Keywords in Portuguese

Keywords

DeCS

Publisher

American Chemical Society

Citation

DOI

ISSN