title: Characterisation of the fumarate hydratase repertoire in Trypanosoma cruzi.
abstract: Nifurtimox and benznidazole represent the only treatments options available targeting Chagas disease, the most important parasitic infection in the Americas. However, use of these is problematic as they are toxic and ineffective against the more severe stages of the disease. In this work, we used a multidisciplinary approach to characterise the fumarases from Trypanosoma cruzi, the causative agent of Chagas Disease. We showed this trypanosome expresses cytosolic and mitochondrial fumarases that via an iron-sulfur cluster mediate the reversible conversion of fumarate to S-malate. Based on sequence, biochemical properties and co-factor binding, both T. cruzi proteins share characteristics with class I fumarases, enzymes found in bacteria and some other protozoa but absent from humans, that possess class II isoforms instead. Gene disruption suggested that although the cytosolic or mitochondrial fumarase activities are individually dispensable their combined activity is essential for parasite viability. Finally, based on the mechanistic differences with the human (host) fumarase, we designed and validated a selective inhibitor targeting the parasite enzyme. This study showed that T. cruzi fumarases should be exploited as targets for the development of new chemotherapeutic interventions against Chagas disease.
authors.affiliation: Laboratório de Cristalografia de Proteínas de Ribeirão Preto, Faculdade de Ciências Farmacêuticas de Ribeirão Preto-Universidade de São Paulo, 14040-903, Ribeirão Preto, SP, Brazil; School of Biological and Chemical Sciences, Queen Mary University of London, Mile End Road, London E1 4NS, United Kingdom.
authors.forename: Ricardo A P
authors.initials: RAP
authors.lastname: de Pádua
authors.affiliation: School of Biological and Chemical Sciences, Queen Mary University of London, Mile End Road, London E1 4NS, United Kingdom.
authors.forename: Ali Martin
authors.initials: AM
authors.lastname: Kia
authors.affiliation: Laboratório de Biofísica Molecular, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto-Universidade de São Paulo, 14040-901, Ribeirão Preto, SP, Brazil.
authors.forename: Antonio J
authors.initials: AJ
authors.lastname: Costa-Filho
authors.affiliation: School of Biological and Chemical Sciences, Queen Mary University of London, Mile End Road, London E1 4NS, United Kingdom. Electronic address: s.r.wilkinson@qmul.ac.uk.
authors.forename: Shane R
authors.initials: SR
authors.lastname: Wilkinson
authors.affiliation: Laboratório de Cristalografia de Proteínas de Ribeirão Preto, Faculdade de Ciências Farmacêuticas de Ribeirão Preto-Universidade de São Paulo, 14040-903, Ribeirão Preto, SP, Brazil. Electronic address: cristy@fcfrp.usp.br.
authors.forename: M Cristina
authors.initials: MC
authors.lastname: Nonato
mesh_terms: D003600:Cytosol / Q000201:enzymology; D055808:Drug Discovery; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005649:Fumarate Hydratase / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000378:metabolism*; D008928:Mitochondria / Q000201:enzymology; D021381:Protein Transport; D014349:Trypanosoma cruzi / Q000166:cytology / Q000201:enzymology*
chemical_list: D004791:Enzyme Inhibitors; D005649:Fumarate Hydratase
keywords: Drug design; Enzyme inhibitor; Gene disruption; Iron–sulfur protein; Tricarboxylic acid cycle; Trypanosoma cruzi