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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: [email protected]
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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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References

REFERENCES

Bermudes, D., Peck, K. R., Afifi, M. A., Beckers, C. J. M. and Joiner, K. A. (1994). Tandemly repeated genes encode nucleoside triphosphate hydrolase isoforms secreted into the parasitophorous vacuole of Toxoplasma gondii. The Journal of Biological Chemistry 269, 29252–29260.Google Scholar
Borges, F. P., de Brum Vieira, P., Wiltuschnig, R. C., Tasca, T., De Carli, G. A. and Bonan, C. D. (2008). Characterization of nucleoside triphosphate diphosphohydrolase activity in Trichomonas gallinae and the influence of penicillin and streptomycin in extracellular nucleotide hydrolysis. FEMS Microbiology Letters 283, 189–195.CrossRefGoogle ScholarPubMed
Burnstock, G. and Verkhratsky, A. (2009). Evolutionary origins of the purinergic signaling system. Acta Physiologica 195, 415–447.CrossRefGoogle Scholar
Coimbra, E. S., Gonçalves-Da-Costa, S. C., Corte-Real, S., Freitas, F. G. R., Durão, A., Souza, C. S. F., Silva Santos, M. I. and Vasconcelos, E. G. (2002). Characterization and cytochemical localization of an ATP diphosphohydrolase from Leishmania amazonensis promastigote. Parasitology 124, 137–143.Google Scholar
Coimbra, E. S., Gonçalves-Da-Costa, S. C., Costa, B. L. S., Giarola, N. L. L., Rezende-Soares, F. A., Fessel, M. R., Ferreira, A. P., Souza, C. S. F., Abreu-Silva, A. L. and Vasconcelos, E. G. (2008). A Leishmania (L.) amazonensis ATP diphosphohydrolase isoform and potato apyrase share epitopes: antigenicity and correlation with disease progression. Parasitology 135, 327–335.CrossRefGoogle ScholarPubMed
Faria-Pinto, P., Meirelles, M. N. L., Lenzi, H. L., Mota, E. M., Penido, M. L. O., Coelho, P. M. Z. and Vasconcelos, E. G. (2004). ATP diphosphohydrolase from Schistosoma mansoni egg: characterization of a new antigen. Parasitology 129, 51–57.CrossRefGoogle ScholarPubMed
Faria-Pinto, P., Meirelles, M. N. L., Lenzi, H. L., Mota, E. M., Penido, M. L. O., Coelho, P. M. Z. and Vasconcelos, E. G. (2006). Cross-immunoreactivity between anti-potato apyrase antibodies and mammalian ATP diphosphohydrolases: potential use of the vegetal protein in experimental schistosomiasis. Memórias do Instituto Oswaldo Cruz 101, 359–363.Google Scholar
Faria-Pinto, P., Rezende-Soares, F. A., Molica, A. M., Montesano, M. A., Marques, M. J., Rocha, M. O. C., Gomes, J. A. S., Enk, M. J., Correa-Oliveira, R., Coelho, P. M. Z., Neto, S. M., Franco, O. L. and Vasconcelos, E. G. (2008). Mapping of the conserved antigenic domains shared between potato apyrase and parasites ATP diphosphohydrolases: potential application in human parasitic diseases. Parasitology 135, 943–953.CrossRefGoogle ScholarPubMed
Gendron, F. P., Benrezzak, O., Krugh, B. W., Kong, Q., Weisman, G. A. and Beaudoin, A. R. (2002). Purine signaling and potential new therapeutic approach: possible outcomes of NTPDase inhibition. Current Drug Targets 3, 229–245.Google Scholar
Gomes-Silva, A., Souza, M. A., Afonso-Cardoso, S. R., Andrade, L. R., Dietze, R., Lemos, E., Belli, A., Favoreto Júnior, S. and Ferreira, M. S. (2008). Serological reactivity of different antigenic preparations of Leishmania (Leishmania) amazonensis and the Leishmania braziliensis complex. Revista da Sociedade Brasileira de Medicina Tropical 41, 135–141.Google Scholar
Gonçalves-Da-Costa, S. C. (2005). A modulação da resistência do hospedeiro por microorganismos. In Dinâmica das doenças infecciosas e Parasitárias (ed. Coura, J. R.), pp. 165–177. Guanabara Koogan, Rio de Janeiro, Brazil.Google Scholar
Guevara-Flores, A., Olvera-Sánchez, S., Gómez-Concha, C., Juarez, O., Esparza-Perusquía, M., Pardo, J. P., Mendoza-Hernández, G., Martinez, F. and Flores-Herrera, O. (2008). 5-p-Fluorosulfonyl benzoyl adenosine inhibits an ecto-ATP diphosphohydrolase in the tegument surface of Taenia crassiceps cysticerci. Molecular and Biochemical Parasitology 162, 123–133.CrossRefGoogle ScholarPubMed
Junqueira-Pedras, M., Orsini, M., Castro, M., Passos, V. M. A. and Rabello, A. (2003). Antibody subclass profile against Leishmania braziliensis and Leishmania amazonensis in the diagnosis and follow-up of mucosal leishmaniasis. Diagnostic Microbiology and Infectious Disease 47, 477–485.Google Scholar
Kedzierski, L., Zhu, Y. and Handman, E. (2006). Leismania vaccines: progress and problems. Parasitology 133, S87–S112.Google Scholar
Kettlun, A. M., Espinosa, V., Garcia, L. and Valenzuela, M. A. (2005). Potato tuber isoapyrases: substrate specificity, affinity labeling, and proteolytic susceptibility. Phytochemistry 66, 975–982.Google Scholar
Korkmaz, B., Attucci, S., Juliano, M. A., Kalupov, T., Jourdan, M. L., Juliano, L. and Gauthier, F. (2008). Measuring elastase, proteinase 3 and cathepsin G activities at the surface of human neutrophils with fluorescence resonance energy transfer substrates. Nature Protocols 3, 991–1000.Google Scholar
Lowry, O. H., Roseborough, N. J., Farr, A. L. and Randall, R. J. (1951). Protein measurement with the Folin phenol reagent. The Journal of Biological Chemistry 193, 265–275.CrossRefGoogle ScholarPubMed
Marques, M. J., Volpini, A. C., Machado-Coelho, G. L. L., Machado-Pinto, J., da Costa, C. A., Mayrink, W., Genaro, O. and Romanha, A. J. (2006). Comparison of polymerase chain reaction with other laboratory methods for the diagnosis of American cutaneous leishmaniasis diagnosis of cutaneous leishmaniasis by polymerase chain reaction. Diagnostic Microbiology and Infectious Disease 54, 37–43.CrossRefGoogle ScholarPubMed
Passos, V. M. A., Fernandes, O., Lacerda, P. A. F., Volpini, A. C., Gontijo, C. M. F., Degrave, W. and Romanha, A. J. (1999). Leishmania (Viannia) braziliensis is the predominant species infecting patients with American cutaneous leishmaniasis in the State of Minas Gerais, Southeast Brazil. Acta Tropica 72, 251–258.CrossRefGoogle Scholar
Peacock, C. S., Seeger, K., Harris, D., Murphy, L., Ruiz, J. C., Quail, M. A., Peters, N., Adlem, E., Tivey, A., Aslett, M., Kerhornou, A., Ivens, A., Fraser, A., Rajandream, M-A., Carver, T., Norbertczak, H., Chillingworth, T., Hance, Z., Jagels, K., Moule, S., Ormond, D., Rutter, S., Squares, R., Whitehead, S., Rabbinowitsch, E., Arrowsmith, C., White, B., Thurston, S., Bringaud, F., Baldauf, S. L., Faulconbridge, A., Jeffares, D., Depledge, D. P., Oyola, S. O., Hilley, J. D., Brito, L. O., Tosi, L. R. O., Barrell, B., Cruz, A. K., Mottram, J. C., Smith, D. F. and Berriman, M. (2007). Comparative genomic analysis of three Leishmania species that cause diverse human disease. Nature Genetics 39, 839–847.CrossRefGoogle ScholarPubMed
Penido, M. L., Resende, D. M., Vianello, M. A., Bordin, F. H. S., Jacinto, A. A., Dias, W. D., Montesano, M. A., Nelson, D. L., Coelho, P. M. Z. and Vasconcelos, E. G. (2007). A new series of schistosomicide drugs, the alkylaminoalkanethiosulfuric acids, partially inhibit the activity of Schistosoma mansoni ATP diphosphohydrolase. European Journal of Pharmacology 570, 10–17.CrossRefGoogle Scholar
Pinheiro, C. M., Martins-Duarte, E. S., Ferraro, R. B., Souza, A. L. F., Gomes, M. T., Lopes, A. H. C. S., Vannier-Santos, M. A., Santos, A. L. S. and Meyer-Fernandes, J. R. (2006). Leishmania amazonensis: biological and biochemical characterization of ecto-nucleoside triphosphate diphosphohydrolase activities. Experimental Parasitology 114, 16–25.CrossRefGoogle ScholarPubMed
Purcell, A. W., McCluskey, J. and Rossjohn, J. (2007). More than one reason to rethink the use of peptides in vaccine design. Nature Reviews. Drug Discovery 6, 404–414.CrossRefGoogle Scholar
Requena, J. M., Alonso, C. and Soto, M. (2000). Evolutionarily conserved proteins as prominent immunogens during Leishmania infections. Parasitology Today 16, 246–250.Google Scholar
Sansom, F. M., Riedmaier, P., Newton, H. J., Dunstone, M. A., Muller, C. E., Stephan, H., Byres, E., Beddoe, T., Rossjohn, J., Cowan, P. J., d'Apice, A. J., Robson, S. C. and Hartland, E. L. (2008). Enzymatic properties of an ecto-nucleoside triphosphate diphosphohydrolase from Legionella pneumophila: substrate specificity and requirement for virulence. The Journal of Biological Chemistry 283, 12909–12918.CrossRefGoogle ScholarPubMed
Smith, D. F., Peacock, C. S. and Cruz, A. K. (2007). Comparative genomics: from genotype to disease phenotype in the leishmaniasis. International Journal for Parasitology 37, 1173–1186.Google Scholar
Taussky, H. M. and Shorr, E. (1953). A microcolorimetric method for the determination of inorganic phosphorus. The Journal of Biological Chemistry 202, 675–685.CrossRefGoogle ScholarPubMed
Vasconcelos, E. G., Faria-Pinto, P., Rezende-Soares, F. A., Penido, M. L., Gonçalves-da-Costa, S. C. and Coelho, P. M. Z. (2009). Potato apyrase: a new tool for parasitic disease research. In Potato II. Fruit, Vegetable and Cereal Science and Biotechnology 3 (Special Issue 1), (ed. Tennant, P. and Benkeblia, N.),p. 80–88. Global Science Books, Ltd., Ikenobe, Japan.Google Scholar