title: Trypanosoma cruzi nucleoside triphosphate diphosphohydrolase 1 (TcNTPDase-1) biochemical characterization, immunolocalization and possible role in host cell adhesion.
abstract: Previous work has suggested that Trypanosoma cruzi diphosphohydrolase 1 (TcNTPDase-1) may be involved in the infection of mammalian cells and serve as a potential target for rational drug design. In this work, we produced recombinant TcNTPDase-1 and evaluated its nucleotidase activity, cellular localization and role in parasite adhesion to mammalian host cells. TcNTPDase-1 was able to utilize a broad range of triphosphate and diphosphate nucleosides. The enzyme's Km for ATP (0.096 mM) suggested a capability to influence the host's ATP-dependent purinergic signaling. The use of specific polyclonal antibodies allowed us to confirm the presence of TcNTPDase-1 at the surface of parasites by confocal and electron microscopy. In addition, electron microscopy revealed that TcNTPDase-1 was also found in the flagellum, flagellum insertion region, kinetoplast, nucleus and intracellular vesicles. The presence of this enzyme in the flagellum insertion region and vesicles suggests that it may have a role in nutrient acquisition, and the widespread distribution of TcNTPDase-1 within the parasite suggests that it may be involved in other biological process. Adhesion assays using anti-TcNTPDase-1 polyclonal antibodies as a blocker or purified recombinant TcNTPDase-1 as a competitor revealed that the enzyme has a role in parasite-host cell adhesion. These data open new frontiers to future studies on this specific parasite-host interaction and other unknown functions of TcNTPDase-1 related to its ubiquitous localization.
authors.affiliation: Departamento de Bioquímica e Biologia Molecular, Universidade Federal de Viçosa, Viçosa, CEP 36570-000 MG, Brazil; Instituto Nacional de Biotecnologia Estrutural e Química Medicinal em Doenças Infecciosas - INBEQMeDI, Brazil.
authors.forename: Christiane
authors.initials: C
authors.lastname: Mariotini-Moura
authors.affiliation: Departamento de Bioquímica e Biologia Molecular, Universidade Federal de Viçosa, Viçosa, CEP 36570-000 MG, Brazil; Instituto Nacional de Biotecnologia Estrutural e Química Medicinal em Doenças Infecciosas - INBEQMeDI, Brazil.
authors.forename: Matheus
authors.initials: M
authors.lastname: Silva e Bastos
authors.affiliation: Departamento de Bioquímica e Biologia Molecular, Universidade Federal de Viçosa, Viçosa, CEP 36570-000 MG, Brazil.
authors.forename: Felipe Freitas
authors.initials: FF
authors.lastname: de Castro
authors.affiliation: Departamento de Bioquímica e Biologia Molecular, Universidade Federal de Viçosa, Viçosa, CEP 36570-000 MG, Brazil.
authors.forename: Mellina Lanna
authors.initials: ML
authors.lastname: Trindade
authors.affiliation: Departamento de Bioquímica e Biologia Molecular, Universidade Federal de Viçosa, Viçosa, CEP 36570-000 MG, Brazil; Instituto Nacional de Biotecnologia Estrutural e Química Medicinal em Doenças Infecciosas - INBEQMeDI, Brazil.
authors.forename: Raphael de Souza
authors.initials: Rde S
authors.lastname: Vasconcellos
authors.affiliation: Departamento de Bioquímica e Biologia Molecular, Universidade Federal de Viçosa, Viçosa, CEP 36570-000 MG, Brazil; Instituto Nacional de Biotecnologia Estrutural e Química Medicinal em Doenças Infecciosas - INBEQMeDI, Brazil.
authors.forename: Myrian Augusta Araújo
authors.initials: MA
authors.lastname: Neves-do-Valle
authors.affiliation: Departamento de Bioquímica e Biologia Molecular, Universidade Federal de Viçosa, Viçosa, CEP 36570-000 MG, Brazil.
authors.forename: Bernardo Pereira
authors.initials: BP
authors.lastname: Moreira
authors.affiliation: Departamento de Bioquímica e Biologia Molecular, Universidade Federal de Viçosa, Viçosa, CEP 36570-000 MG, Brazil; Instituto Nacional de Biotecnologia Estrutural e Química Medicinal em Doenças Infecciosas - INBEQMeDI, Brazil.
authors.forename: Ramon de Freitas
authors.initials: Rde F
authors.lastname: Santos
authors.affiliation: Departamento de Bioquímica e Biologia Molecular, Universidade Federal de Viçosa, Viçosa, CEP 36570-000 MG, Brazil; Instituto Nacional de Biotecnologia Estrutural e Química Medicinal em Doenças Infecciosas - INBEQMeDI, Brazil.
authors.forename: Claudia Miranda
authors.initials: CM
authors.lastname: de Oliveira
authors.affiliation: Departamento de Bioquímica e Biologia Molecular, Universidade Federal de Viçosa, Viçosa, CEP 36570-000 MG, Brazil.
authors.forename: Luana Celina Seraphim
authors.initials: LC
authors.lastname: Cunha
authors.affiliation: Departamento de Bioquímica e Biologia Molecular, Universidade Federal de Viçosa, Viçosa, CEP 36570-000 MG, Brazil.
authors.forename: Xênia Macedo
authors.initials: XM
authors.lastname: Souto
authors.affiliation: Departamento de Bioquímica e Biologia Molecular, Universidade Federal de Viçosa, Viçosa, CEP 36570-000 MG, Brazil.
authors.forename: Gustavo Costa
authors.initials: GC
authors.lastname: Bressan
authors.affiliation: Departamento de Veterinária, Universidade Federal de Viçosa, Viçosa, CEP 36570-000 MG, Brazil.
authors.forename: Abelardo
authors.initials: A
authors.lastname: Silva-Júnior
authors.affiliation: Departamento de Biologia Celular e Molecular e Bioagentes Patogênicos, Faculdade De Medicina de Ribeirao Preto, Universidade de São Paulo, Ribeirão Preto, CEP 14090-900 SP, Brazil.
authors.forename: Munira Muhammad Abdel
authors.initials: MM
authors.lastname: Baqui
authors.affiliation: Núcleo de Pesquisa em Ciências Biológicas - NUPEB, Universidade Federal de Ouro Preto, Ouro Preto, CEP 35400-000 MG, Brazil.
authors.forename: Maria Terezinha
authors.initials: MT
authors.lastname: Bahia
authors.affiliation: Departamento de Bioquímica e Biologia Molecular, Universidade Federal de Viçosa, Viçosa, CEP 36570-000 MG, Brazil.
authors.forename: Márcia Rogéria
authors.initials: MR
authors.lastname: de Almeida
authors.affiliation: Instituto de Bioquímica Médica, Universidade Federal do Rio de Janeiro, Rio de Janeiro, CEP 21941-590 RJ, Brazil; Instituto Nacional de Biologia Estrutural e Bioimagem, IMBEB, Brazil.
authors.forename: José Roberto
authors.initials: JR
authors.lastname: Meyer-Fernandes
authors.affiliation: Departamento de Bioquímica e Biologia Molecular, Universidade Federal de Viçosa, Viçosa, CEP 36570-000 MG, Brazil; Instituto Nacional de Biotecnologia Estrutural e Química Medicinal em Doenças Infecciosas - INBEQMeDI, Brazil. Electronic address: jufietto@ufv.br.
authors.forename: Juliana Lopes Rangel
authors.initials: JL
authors.lastname: Fietto
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000737:chemistry / Q000502:physiology*; D001081:Apyrase / Q000737:chemistry / Q000502:physiology*; D015153:Blotting, Western; D002448:Cell Adhesion / Q000502:physiology*; D006790:Host-Parasite Interactions / Q000502:physiology*; D007150:Immunohistochemistry; D014349:Trypanosoma cruzi / Q000201:enzymology*
chemical_list: D015703:Antigens, CD; D001081:Apyrase; C067869:CD39 antigen
keywords: Adhesion; Immunolocalization; Nucleoside triphosphate diphosphohydrolase; Recombinant protein; Trypanosoma cruzi