Please use this identifier to cite or link to this item: https://www.arca.fiocruz.br/handle/icict/23183
Title: Rapid Chagas Disease Drug Target Discovery Using Directed Evolution in Drug-Sensitive Yeast
Authors: Ottilie, Sabine
Goldgof, Gregory M.
Calvet, Claudia Magalhaes
Jennings, Gareth K.
LaMonte, Greg
Schenken, Jake
Vigil, Edgar
Kumar, Prianka
McCall, Laura-Isobel
Lopes, Eduardo Soares Constantino
Gunawan, Felicia
Yang, Jennifer
Suzuki, Yo
Siqueira Neto, Jair L.
McKerrow, James H.
Amaro, Rommie E.
Podust, Larissa M.
Durrant, Jacob D.
Winzeler, Elizabeth A.
Affilliation: 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.
Keywords: Chagas Disease
Trypanosoma cruzi
ergosterol
Cyp51
ERG11
S. cerevisiae
directed evolution
whole genome sequencing
keywords: Doença de Chagas
Trypanosoma cruzi
evolução dirigida
sequenciação completa do genoma
DeCS: Família 51 do Citocromo P450
Ergosterol
Issue Date: 2017
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
Copyright: restricted access
Appears in Collections:IOC - Artigos de Periódicos

Files in This Item:
File Description SizeFormat 
claudia_calvet_etal_IOC_2017.pdf2.09 MBAdobe PDF    Request a copy


FacebookTwitterDeliciousLinkedInGoogle BookmarksBibTex Format mendeley Endnote DiggMySpace

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.