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Toxoplasma gondii: molecular cloning and characterization of a nitric oxide synthase-like protein.
Gutierrez-Escobar, Andrés J; Arenas, Aylan Farid; Villoria-Guerrero, Yanet; Padilla-Londoño, Jonathan M; Gómez-Marin, Jorge Enrique.
Afiliação
  • Gutierrez-Escobar AJ; Grupo de Parasitología Molecular (GEPAMOL), Centro de Investigaciones Biomédicas, Facultad de Ciencias de la Salud, Universidad del Quindío, Armenia, Colombia.
Exp Parasitol ; 119(3): 358-63, 2008 Jul.
Article em En | MEDLINE | ID: mdl-18439581
Toxoplasma gondii has a nitrite production and a putative nitric oxide synthase (NOS) motif genomic sequence. In order to demonstrate that this sequence is functional and could be involved in the metabolism of l-arginine derivatives, we constructed a baculovirus carrying the previously identified Toxoplasma NOS-like DNA sequence. The recombinant protein was expressed into insect Sf9 cells and his activity was tested in serial microplate colorimetric assays. The protein produced 21 nmol/min/ml nitrites per microgram of protein and followed Michaelis-Menten kinetics, with a K(m) for L-arginine of 2.3mM. Furthermore, the optimal pH, temperature and incubation time for the recombinant Toxoplasma NOS-like protein were established. Toxoplasma NOS runs as a band of 11.6 kDa on tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Our results indicate that the recombinant protein derived from the putative genomic sequence, at the chromosome 1b of T. gondii, is able to produce nitrites from L-arginine as substrate.
Assuntos

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Toxoplasma / Óxido Nítrico Sintase Limite: Animals Idioma: En Revista: Exp Parasitol Ano de publicação: 2008 Tipo de documento: Article País de afiliação: Colômbia País de publicação: Estados Unidos

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Toxoplasma / Óxido Nítrico Sintase Limite: Animals Idioma: En Revista: Exp Parasitol Ano de publicação: 2008 Tipo de documento: Article País de afiliação: Colômbia País de publicação: Estados Unidos