Cytochemical localization of ATP diphosphohydrolase from Leishmania (Viannia) braziliensis promastigotes and identification of an antigenic and catalytically active isoform

Cytochemical localization of ATP diphosphohydrolase from Leishmania (Viannia) braziliensis promastigotes and identification of an antigenic and catalytically active isoform

Autor Rezende-Soares, F. A. Google Scholar
Carvalho-Campos, C. Google Scholar
Marques, M. J. Google Scholar
Porcino, G. N. Google Scholar
Giarola, N. L. L. Google Scholar
Costa, B. L. S. Google Scholar
Taunay-Rodrigues, A. Google Scholar
Faria-Pinto, P. Google Scholar
Souza, M. A. Google Scholar
Diniz, V. A. Google Scholar
Corte-Real, S. Google Scholar
Juliano, M. A. Autor UNIFESP Google Scholar
Juliano, L. Autor UNIFESP Google Scholar
Vasconcelos, E. G. Google Scholar
Instituição Univ Fed Juiz de Fora
Univ Fed Alfenas
Universidade Federal de Uberlândia (UFU)
Fiocruz MS
Universidade Federal de São Paulo (UNIFESP)
Resumo 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.
Palavra-chave ATP diphosphohydrolase
potato apyrase
NDPase
GDPase
ecto-enzyme
Leishmania (Viannia) braziliensis
promastigote
American cutaneous leishmaniasis
Idioma Inglês
Financiador Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG)
Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
UFJF/MG
PIBIC
PROBIC
BIC/UFJF
Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Número do financiamento FAPEMIG: CBB-1859/06
CNPq: 474158/2003-9
Data de publicação 2010-04-01
Publicado em Parasitology. New York: Cambridge Univ Press, v. 137, n. 5, p. 773-783, 2010.
ISSN 0031-1820 (Sherpa/Romeo, fator de impacto)
Publicador Cambridge Univ Press
Extensão 773-783
Fonte http://dx.doi.org/10.1017/S0031182009991661
Direito de acesso Acesso restrito
Tipo Artigo
Web of Science WOS:000278283400002
Endereço permanente http://repositorio.unifesp.br/handle/11600/32439

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