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

Published online by Cambridge University Press:  07 December 2009

F. A. REZENDE-SOARES ,
C. CARVALHO-CAMPOS ,
M. J. MARQUES ,
G. N. PORCINO ,
N. L. L. GIAROLA ,
B. L. S. COSTA ,
A. TAUNAY-RODRIGUES ,
P. FARIA-PINTO ,
M. A. SOUZA  and
V. A. DINIZ
...Show all authors
F. A. REZENDE-SOARES
Affiliation:
Departamento de Bioquímica/Pós-Graduação em Imunologia, Genética e Biotecnologia/ICB, Universidade Federal de Juiz de Fora, Campus Universitário, Bairro Cidade Universitária, Juiz de Fora, MG, Brazil
C. CARVALHO-CAMPOS
Affiliation:
Departamento de Bioquímica/Pós-Graduação em Imunologia, Genética e Biotecnologia/ICB, Universidade Federal de Juiz de Fora, Campus Universitário, Bairro Cidade Universitária, Juiz de Fora, MG, Brazil
M. J. MARQUES
Affiliation:
Departamento de Bioquímica/Pós-Graduação em Imunologia, Genética e Biotecnologia/ICB, Universidade Federal de Juiz de Fora, Campus Universitário, Bairro Cidade Universitária, Juiz de Fora, MG, Brazil Departamento de Ciências Biológicas, Universidade Federal de Alfenas, Alfenas, MG, Brazil
G. N. PORCINO
Affiliation:
Departamento de Bioquímica/Pós-Graduação em Imunologia, Genética e Biotecnologia/ICB, Universidade Federal de Juiz de Fora, Campus Universitário, Bairro Cidade Universitária, Juiz de Fora, MG, Brazil
N. L. L. GIAROLA
Affiliation:
Departamento de Bioquímica/Pós-Graduação em Imunologia, Genética e Biotecnologia/ICB, Universidade Federal de Juiz de Fora, Campus Universitário, Bairro Cidade Universitária, Juiz de Fora, MG, Brazil
B. L. S. COSTA
Affiliation:
Departamento de Bioquímica/Pós-Graduação em Imunologia, Genética e Biotecnologia/ICB, Universidade Federal de Juiz de Fora, Campus Universitário, Bairro Cidade Universitária, Juiz de Fora, MG, Brazil
A. TAUNAY-RODRIGUES
Affiliation:
Departamento de Bioquímica/Pós-Graduação em Imunologia, Genética e Biotecnologia/ICB, Universidade Federal de Juiz de Fora, Campus Universitário, Bairro Cidade Universitária, Juiz de Fora, MG, Brazil
P. FARIA-PINTO
Affiliation:
Departamento de Bioquímica/Pós-Graduação em Imunologia, Genética e Biotecnologia/ICB, Universidade Federal de Juiz de Fora, Campus Universitário, Bairro Cidade Universitária, Juiz de Fora, MG, Brazil
M. A. SOUZA
Affiliation:
Departamento de Bioquímica/Pós-Graduação em Imunologia, Genética e Biotecnologia/ICB, Universidade Federal de Juiz de Fora, Campus Universitário, Bairro Cidade Universitária, Juiz de Fora, MG, Brazil Laboratório de Imunologia, Instituto de Ciências Biomédicas, Universidade Federal de Uberlândia, Uberlândia, MG, Brazil
V. A. DINIZ
Affiliation:
Laboratório de Biologia Estrutural, Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro, RJ, Brazil
S. CORTE-REAL
Affiliation:
Laboratório de Biologia Estrutural, Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro, RJ, Brazil
M. A. JULIANO
Affiliation:
Departamento de Biofísica, Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, SP, Brazil
L. JULIANO
Affiliation:
Departamento de Biofísica, Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, SP, Brazil
E. G. VASCONCELOS*
Affiliation:
Departamento de Bioquímica/Pós-Graduação em Imunologia, Genética e Biotecnologia/ICB, Universidade Federal de Juiz de Fora, Campus Universitário, Bairro Cidade Universitária, Juiz de Fora, MG, Brazil
*
*Corresponding author: Departamento de Bioquímica, ICB, Universidade Federal de Juiz de Fora, Campus Universitário, Bairro Cidade Universitária, 36036-330, Juiz de Fora, MG, Brazil. Tel: +55 32 2102 3217. Fax: +55 32 2102 3216. E-mail: eveline.vasconcelos@ufjf.edu.br
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Summary

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.

Keywords

ATP diphosphohydrolasepotato apyraseNDPaseGDPaseecto-enzymeLeishmania (Viannia) braziliensispromastigoteAmerican cutaneous leishmaniasis
Type
Research Article
Information
Parasitology , Volume 137 , Issue 5 , April 2010 , pp. 773 - 783
Copyright
Copyright © Cambridge University Press 2009

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