title: Differential Gel Electrophoresis (DIGE) Evaluation of Naphthoimidazoles Mode of Action: A Study in Trypanosoma cruzi Bloodstream Trypomastigotes.
abstract: The obligate intracellular protozoan Trypanosoma cruzi is the causative agent of Chagas disease, a neglected illness affecting millions of people in Latin America that recently entered non-endemic countries through immigration, as a consequence of globalization. The chemotherapy for this disease is based mainly on benznidazole and nifurtimox, which are very efficient nitroderivatives against the acute stage but present limited efficacy during the chronic phase. Our group has been studying the trypanocidal effects of naturally occurring quinones and their derivatives, and naphthoimidazoles derived from β-lapachone N1, N2 and N3 were the most active. To assess the molecular mechanisms of action of these compounds, we applied proteomic techniques to analyze treated bloodstream trypomastigotes, which are the clinically relevant stage of the parasite.
  
  
  
  The approach consisted of quantification by 2D-DIGE followed by MALDI-TOF/TOF protein identification. A total of 61 differentially abundant protein spots were detected when comparing the control with each N1, N2 or N3 treatment, for 34 identified spots. Among the differentially abundant proteins were activated protein kinase C receptor, tubulin isoforms, asparagine synthetase, arginine kinase, elongation factor 2, enolase, guanine deaminase, heat shock proteins, hypothetical proteins, paraflagellar rod components, RAB GDP dissociation inhibitor, succinyl-CoA ligase, ATP synthase subunit B and methionine sulfoxide reductase.
  
  
  
  Our results point to different modes of action for N1, N2 and N3, which indicate a great variety of metabolic pathways involved and allow for novel perspectives on the development of trypanocidal agents.
authors.affiliation: Laboratório de Toxinologia, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Rio de Janeiro, Brazil.
authors.forename: Giselle Villa Flor
authors.initials: GV
authors.lastname: Brunoro
authors.affiliation: Departamento de Bioquímica e Imunologia, Faculdade de Medicina de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, Brazil.
authors.forename: Vitor Marcel
authors.initials: VM
authors.lastname: Faça
authors.affiliation: Laboratório de Toxinologia, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Rio de Janeiro, Brazil.|Laboratório de Biologia Celular, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Rio de Janeiro, Brazil.
authors.forename: Marcelle Almeida
authors.initials: MA
authors.lastname: Caminha
authors.affiliation: Laboratório de Toxinologia, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Rio de Janeiro, Brazil.
authors.forename: André Teixeira da Silva
authors.initials: AT
authors.lastname: Ferreira
authors.affiliation: Laboratório de Toxinologia, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Rio de Janeiro, Brazil.
authors.forename: Monique
authors.initials: M
authors.lastname: Trugilho
authors.affiliation: Núcleo de Pesquisas em Produtos Naturais, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.
authors.forename: Kelly Cristina Gallan
authors.initials: KC
authors.lastname: de Moura
authors.affiliation: Laboratório de Toxinologia, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Rio de Janeiro, Brazil.
authors.forename: Jonas
authors.initials: J
authors.lastname: Perales
authors.affiliation: Laboratório de Toxinologia, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Rio de Janeiro, Brazil.
authors.forename: Richard Hemmi
authors.initials: RH
authors.lastname: Valente
authors.affiliation: Laboratório de Biologia Celular, Instituto Oswaldo Cruz, Fundação Oswaldo Cruz, Rio de Janeiro, Brazil.
authors.forename: Rubem Figueiredo Sadok
authors.initials: RF
authors.lastname: Menna-Barreto
mesh_terms: D000818:Animals; D014355:Chagas Disease / Q000188:drug therapy; D015180:Electrophoresis, Gel, Two-Dimensional; D051379:Mice; D009285:Naphthoquinones / Q000494:pharmacology*; D009547:Nifurtimox / Q000494:pharmacology; D009593:Nitroimidazoles / Q000494:pharmacology; D040901:Proteomics; D015800:Protozoan Proteins / Q000032:analysis*; D014344:Trypanocidal Agents / Q000494:pharmacology*; D014349:Trypanosoma cruzi / Q000187:drug effects*
chemical_list: D009285:Naphthoquinones; D009593:Nitroimidazoles; D015800:Protozoan Proteins; D014344:Trypanocidal Agents; C014638:beta-lapachone; D009547:Nifurtimox; C009999:benzonidazole