title: Expression of glyceraldehyde 3-phosphate dehydrogenase is enhanced in Leishmania spp naturally resistant to nitric oxide.
abstract: Leishmania spp are the causative agents of a spectrum of diseases termed leishmaniasis that affect mammals, including humans and dogs. Although reactive nitrogen species are employed in the control of parasitism by the immune system, it is known that Leishmania can withstand this oxidative stress. As the mechanism by which these species are resistant to nitric oxide (NO) is poorly understood, the main objective of this study was to evaluate the expression of glyceraldehyde 3-phosphate dehydrogenase (GAPDH) in Leishmania amazonensis and Leishmania chagasi promastigotes showing natural resistance to NO. GAPDH transcript levels were quantified by real-time polymerase chain reaction amplification, and GAPDH activity (assessed by levels of NADH oxidation) was measured by spectrophotometry. The level of nitration in total protein was assessed by immunoblotting. The results demonstrated an increase in GAPDH expression in resistant isolates of both species compared to susceptible isolates. The increase in GAPDH expression led to an increase in the activity of GAPDH in L. amazonensis human isolates resistant to NO. The pattern of protein nitration did not differ between sensitive and resistant isolates. Our results suggest that changes in expression of GAPDH may be responsible, at least in part, to natural resistance to NO found in human and canine Leishmania spp.
authors.affiliation: Departamento de Fisiologia, Universidade Federal de Sergipe, São Cristóvão, SE, Brasil.
authors.forename: M C
authors.initials: MC
authors.lastname: Rios
authors.affiliation: Departamento de Morfologia, Universidade Federal de Sergipe, São Cristóvão, SE, Brasil.
authors.forename: W R T
authors.initials: WR
authors.lastname: Silva
authors.affiliation: Departamento de Fisiologia, Universidade Federal de Sergipe, São Cristóvão, SE, Brasil.
authors.forename: A F
authors.initials: AF
authors.lastname: Azevedo
authors.affiliation: Departamento de Medicina, Universidade Federal de Sergipe, Aracaju, SE, Brasil.
authors.forename: P L
authors.initials: PL
authors.lastname: Santos
authors.affiliation: Departamento de Farmacologia, Universidade de São Paulo, São Paulo, SP, Brasil.
authors.forename: S A
authors.initials: SA
authors.lastname: Teixeira
authors.affiliation: Departamento de Farmacologia, Universidade de São Paulo, São Paulo, SP, Brasil.
authors.forename: M N
authors.initials: MN
authors.lastname: Muscará
authors.affiliation: Departamento de Fisiologia, Universidade Federal de Sergipe, São Cristóvão, SE, Brasil.
authors.forename: S M
authors.initials: SM
authors.lastname: Thomazzi
authors.affiliation: Departamento de Medicina, Universidade Federal de Sergipe, Aracaju, SE, Brasil.
authors.forename: R P
authors.initials: RP
authors.lastname: Almeida
authors.affiliation: Departamento de Fisiologia, Universidade Federal de Sergipe, São Cristóvão, SE, Brasil.
authors.forename: R P M
authors.initials: RP
authors.lastname: Fernandes
authors.affiliation: Departamento de Morfologia, Universidade Federal de Sergipe, São Cristóvão, SE, Brasil scher@ufs.br.
authors.forename: R
authors.initials: R
authors.lastname: Scher
mesh_terms: D003470:Culture Media; D004351:Drug Resistance; D015870:Gene Expression / Q000187:drug effects*; D024601:Glyceraldehyde 3-Phosphate Dehydrogenase (NADP+) / Q000235:genetics* / Q000378:metabolism; D007891:Leishmania / Q000187:drug effects / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D018314:Leishmania infantum / Q000187:drug effects / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D008018:Life Cycle Stages / Q000187:drug effects*; D009243:NAD / Q000378:metabolism; D009569:Nitric Oxide / Q000378:metabolism / Q000494:pharmacology*; D010084:Oxidation-Reduction; D015800:Protozoan Proteins / Q000235:genetics* / Q000378:metabolism; D012977:Sodium Nitrite / Q000737:chemistry / Q000494:pharmacology
chemical_list: D003470:Culture Media; D015800:Protozoan Proteins; D009243:NAD; D009569:Nitric Oxide; D024601:Glyceraldehyde 3-Phosphate Dehydrogenase (NADP+); D012977:Sodium Nitrite