Navegando por Palavras-chave "potato apyrase"

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    Cytochemical localization of ATP diphosphohydrolase from Leishmania (Viannia) braziliensis promastigotes and identification of an antigenic and catalytically active isoform
    (Cambridge Univ Press, 2010-04-01) Rezende-Soares, F. A.; Carvalho-Campos, C.; Marques, M. J.; Porcino, G. N.; Giarola, N. L. L.; Costa, B. L. S.; Taunay-Rodrigues, A.; Faria-Pinto, P.; Souza, M. A.; Diniz, V. A.; Corte-Real, S.; Juliano, M. A. [UNIFESP]; Juliano, L. [UNIFESP]; Vasconcelos, E. G.; Univ Fed Juiz de Fora; Univ Fed Alfenas; Universidade Federal de Uberlândia (UFU); Fiocruz MS; Universidade Federal de São Paulo (UNIFESP)
    An ATP diphosphohydrolase (EC 3.6.1.5) activity was identified in a Leishmania (Viannia) braziliensis promastigotes preparation (Lb). Ultrastructural cytochemical microscopy showed this protein on the parasite surface and also stained a possible similar protein at the mitochondrial membrane. Isolation of an active ATP diphosphohydrolase isoform from Lb was obtained by cross-immunoreactivity with polyclonal anti-potato apyrase antibodies. These antibodies, immobilized on Protein A-Sepharose, immunoprecipitated a polypeptide of approximately 48 kDa and, in lower amount, a polypeptide of approximately 43 kDa, and depleted 83% ATPase and 87% of the ADPase activities from detergent-homogenized Lb. Potato apyrase was recognized in Western blots by IgG antibody from American cutaneous leishmaniasis (ACL) patients, suggesting that the parasite and vegetable proteins share antigenic conserved epitopes. Significant IgG seropositivity in serum samples diluted 1 :50 from ACL patients (n=20) for Lb (65%) and potato apyrase (90%) was observed by ELISA technique. Significant IgG antibody reactivity was also observed against synthetic peptides belonging to a conserved domain from L. braziliensis NDPase (80% seropositivity) and its potato apyrase counterpart (50% seropositivity), in accordance with the existence of shared antigenic epitopes and demonstrating that in leishmaniasis infection the domain r82-103 from L. braziliensis NDPase is a target for the human immune response.
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    Immunostimulatory property of a synthetic peptide belonging to the soluble ATP diphosphohydro-lase isoform (SmATPDase 2) and immunolocalisation of this protein in the Schistosoma mansoni egg
    (Instituto Oswaldo Cruz, Ministério da Saúde, 2011-11-01) Mendes, Rita Gabriela Pedrosa Ribeiro; Gusmão, Michélia Antônia do Nascimento [UNIFESP]; Maia, Ana Carolina Ribeiro Gomes; Detoni, Michelle de Lima; Porcino, Gabriane Nascimento; Soares, Thais Vieira; Juliano, Maria Aparecida [UNIFESP]; Juliano, Luiz [UNIFESP]; Coelho, Paulo Marcos Zech; Lenzi, Henrique Leonel; Faria-Pinto, Priscila; Vasconcelos, Eveline Gomes; Universidade Federal de Juiz de Fora Instituto de Ciências Biológicas Departamento de Bioquímica; Universidade Federal de São Paulo (UNIFESP); Instituto de Pesquisas René Rachou-Fiocruz; Instituto Oswaldo Cruz-Fiocruz Laboratório de Patologia
    A peptide (SmB2LJ; r175-194) that belongs to a conserved domain from Schistosoma mansoni SmATPDase 2 and is shared with potato apyrase, as predicted by in silico analysis as antigenic, was synthesised and its immunostimulatory property was analysed. When inoculated in BALB/c mice, this peptide induced high levels of SmB2LJ-specific IgG1 and IgG2a subtypes, as detected by enzyme linked immunosorbent assay. In addition, dot blots were found to be positive for immune sera against potato apyrase and SmB2LJ. These results suggest that the conserved domain r175-194 from the S. mansoni SmATPDase 2 is antigenic. Western blots were performed and the anti-SmB2LJ antibody recognised in adult worm (soluble worm antigen preparation) or soluble egg antigen antigenic preparations two bands of approximately 63 and 55 kDa, molecular masses similar to those predicted for adult worm SmATPDase 2. This finding strongly suggests the expression of this same isoform in S. mansoni eggs. To assess localisation of SmATPDase 2, confocal fluorescence microscopy was performed using cryostat sections of infected mouse liver and polyclonal antiserum against SmB2LJ. Positive reactions were identified on the external surface from the miracidium in von Lichtenberg's envelope and, in the outer side of the egg-shell, showing that this soluble isoform is secreted from the S. mansoni eggs.