Hostname: page-component-76fb5796d-9pm4c Total loading time: 0 Render date: 2024-04-25T08:47:07.828Z Has data issue: false hasContentIssue false
  • English
  • Français

CrATP interferes in the promastigote-macrophage interaction in Leishmania amazonensis infection

Published online by Cambridge University Press:  17 June 2011

V. ENNES-VIDAL ,
R. O. S. CASTRO ,
C. BRITTO ,
H. BARRABIN ,
C. M. D'AVILA-LEVY  and
O. C. MOREIRA
V. ENNES-VIDAL
Affiliation:
Laboratório de Biologia Molecular e Doenças Endêmicas, Instituto Oswaldo Cruz/FIOCRUZ, Rio de Janeiro, RJ, Brazil
R. O. S. CASTRO
Affiliation:
Instituto Federal de Educação, Ciência e Tecnologia do Rio de Janeiro, Rio de Janeiro, RJ, Brazil
C. BRITTO
Affiliation:
Laboratório de Biologia Molecular e Doenças Endêmicas, Instituto Oswaldo Cruz/FIOCRUZ, Rio de Janeiro, RJ, Brazil
H. BARRABIN
Affiliation:
Laboratório de Transporte Iônico em Membranas Biológicas, Instituto de Bioquímica Médica, Universidade Federal do Rio de Janeiro, RJ, Brazil
C. M. D'AVILA-LEVY
Affiliation:
Laboratório de Biologia Molecular e Doenças Endêmicas, Instituto Oswaldo Cruz/FIOCRUZ, Rio de Janeiro, RJ, Brazil
O. C. MOREIRA*
Affiliation:
Laboratório de Biologia Molecular e Doenças Endêmicas, Instituto Oswaldo Cruz/FIOCRUZ, Rio de Janeiro, RJ, Brazil Instituto Federal de Educação, Ciência e Tecnologia do Rio de Janeiro, Rio de Janeiro, RJ, Brazil
*
*Corresponding author: Instituto Oswaldo Cruz/FIOCRUZ, CEP: 21040-360, Rio de Janeiro, RJ, Brazil. Tel: +55 21 3865 8153. Fax: +55 21 2590-3495. E-mail: otacilio@ioc.fiocruz.br
Get access Rights & Permissions [Opens in a new window]

Summary

Recent have shown the relationship between Ecto-Nucleoside-Triphosphate-Diphosphohydrolases (Ecto-NTPDases or ecto-nucleotidases) and virulence and infectivity in trypanosomatids. In this work, the inhibition of the ecto-ATPase activities and promastigote growth of Leishmania amazonensis by CrATP was characterized. Furthermore, this compound was used to investigate the role of ecto-nucleotidase in the interaction of L. amazonensis with resident peritoneal macrophages obtained from BALB/c mice. CrATP partially inhibits the ecto-ATPase activity, presenting Ki values of 575·7±199·1 and 383·5±79·0 μm, in the presence or absence of 5 mm MgCl2, respectively. The apparent Kms for ATP (2·9±0·5 mm to Mg2+-dependent ecto-ATPase and 0·4±0·2 mm to Mg2+-independent ecto-ATPase activities) are not significantly altered by CrATP, suggesting a reversible non-competitive inhibition of both enzymes. When CrATP was added to the cultivation medium at 500 μm, it drastically inhibited the cellular growth. The interaction of promastigote forms of L. amazonensis with BALB/c peritoneal macrophages is strongly affected by CrATP. When the parasites were treated with 500 μm CrATP before interacting with macrophages, the adhesion and endocytic indices were strongly reduced to 53·0±14·8% and 39·8±1·1%, respectively. These results indicate that ecto-nucleotidase plays an important role in the infection process caused by Leishmania amazonensis.

Keywords

ecto-NTPDaseCrATPLeishmania amazonensisinfectivity
Type
Research Article
Information
Parasitology , Volume 138 , Issue 8 , July 2011 , pp. 960 - 968
Copyright
Copyright © Cambridge University Press 2011

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

REFERENCES

Barral, A., Pedral-Sampaio, D., Grimaldi Junior, G., Momen, H., McMahon-Pratt, D., Ribeiro de Jesus, A., Almeida, R., Badaro, M., Barral-Neto, M., Carvalho, E. M. and Johnson, W. D. Jr. (1991). Leishmaniasis in Bahia, Brazil: evidence that Leishmania amazonensis produces a wide spectrum of clinical disease. The American Journal of Tropical Medicine and Hygiene 44, 536–546.CrossRefGoogle ScholarPubMed
Barros, F. S., De Menezes, L. F., Pinheiro, A. A., Silva, E. F., Lopes, A. H., De Souza, W. and Meyer-Fernandes, J. R. (2000). Ectonucleotide diphosphohydrolase activities in Entamoeba histolytica. Archives of Biochemistry and Biophysics 375, 304–314.CrossRefGoogle ScholarPubMed
Bernardes, C. F., Meyer-Fernandes, J. R., Saad-Nehme, J., Vannier-Santos, M. A., Peres-Sampaio, C. E. and Vercesi, A. E. (2000). Effects of 4,49-diisothyocyanatostilbene-2,29-disulfonic acid on Trypanosoma cruzi proliferation and Ca(2+) homeostasis. International Journal of Biochemistry and Cell Biology 32, 519–527.CrossRefGoogle Scholar
Berrêdo-Pinho, M., Peres-Sampaio, C. E., Chrispim, P. P., Belmont-Firpo, R., Lemos, A. P., Martiny, A., Vannier-Santos, M. A. and Meyer-Fernandes, J. R. (2001). A Mg-dependent ecto-ATPase in Leishmania amazonensis and its possible role in adenosine acquisition and virulence. Archives of Biochemistry and Biophysics 391, 16–24.CrossRefGoogle ScholarPubMed
Bisaggio, D. F., Peres-Sampaio, C. E., Meyer-Fernandes, J. R. and Souto-Padron, T. (2003). Ecto-ATPase activity on the surface of Trypanosoma cruzi and its possible role in the parasite-host cell interaction. Parasitology Research 91, 273–282.CrossRefGoogle ScholarPubMed
Collopy-Junior, I., Esteves, F. F., Nimrichter, L., Rodrigues, M. L., Alviano, C. S. and Meyer-Fernandes, J. R. (2006). An ectophosphatase activity in Cryptococcus neoformans. FEMS Yeast Research 6, 1010–1017.CrossRefGoogle ScholarPubMed
Convit, J., Pinardi, M. E. and Rondón, A. J. (1972). Diffuse cutaneous leishmaniasis: a disease due to an immunological defect in the host. Transactions of the Royal Society of Tropical Medicine and Hygiene 66, 603–610.CrossRefGoogle Scholar
De Jesus, J. B., de Sá Pinheiro, A. A., Lopes, A. H. and Meyer-Fernandes, J. R. (2002). An ectonucleotide ATP-diphosphohydrolase activity in Trichomonas vaginalis stimulated by galactose and its possible role in virulence. Zeitschrift der Naturforschung C 57, 890–896.CrossRefGoogle ScholarPubMed
De Pierre, J. W. and Karnovsky, M. L. (1973). Plasma membranes of mammalian cells: a review of methods for their characterization and isolation. Journal of Cell Biology 56, 275–303.CrossRefGoogle Scholar
De Pierre, J. W. and Karnovsky, M. L. (1974). Ecto-enzymes of the guinea pig polymorphonuclear leukocyte. Evidence for an ecto-adenosine monophosphatase, adenosine triphosphatase, and p-nitrophenyl phosphates. Journal of Biological Chemistry 249, 7111–7120.CrossRefGoogle ScholarPubMed
DePamphilis, M. L. and Cleland, W. W. (1973). Preparation and properties of chromium(III)-nucleotide complexes for use in the study of enzyme mechanisms. Biochemistry 12, 3714–3724.CrossRefGoogle Scholar
De Souza, M. C., De Assis, E. A., Gomes, R. S., Marques da Silva, E. A., Melo, M. N., Fietto, J. L. and Afonso, L. C. (2010). The influence of ecto-nucleotidases on Leishmania amazonensis infection and immune response in C57B/6 mice. Acta Tropica 115, 262–269.CrossRefGoogle ScholarPubMed
Dutra, P. M., Dias, F. A., Santos, M. A., Rodrigues, C. O., Romeiro, A., Attias, M., De Souza, W., Lopes, A. H. and Meyer-Fernandes, J. R. (2001). Secreted phosphatase activities in trypanosomatid parasites of plants modulated by platelet-activating factor. Phytopathology 91, 408–414.CrossRefGoogle ScholarPubMed
Einholm, A. P., Vilsen, B. and Andersen, J. P. (2004). Importance of transmembrane segment M1 of the sarcoplasmic reticulum Ca2+-ATPase in Ca2+ occlusion and phosphoenzyme processing. Journal of Biological Chemistry 279, 15888–15896.CrossRefGoogle ScholarPubMed
Furuya, T., Zhong, L., Meyer-Fernandes, J. R., Lu, H. G., Moreno, S. N. and Docampo, R. (1998). Ecto-protein tyrosine phosphatase activity in Trypanosoma cruzi infective stages. Molecular and Biochemical Parasitology 92, 339–348.CrossRefGoogle ScholarPubMed
Hopfe, M. and Henrich, B. (2008). OppA, the ecto-ATPase of Mycoplasma hominis induces ATP release and cell death in HeLa cells. BMC Microbiology 4, 8–55.Google Scholar
Jesus, J. B., Lopes, A. H. and Meyer-Fernandes, J. R. (2002). Characterization of an ecto-ATPase of Tritrichomonas foetus. Veterinary Parasitology 103, 29–42.CrossRefGoogle ScholarPubMed
Kiffer-Moreira, T., Fernandes Sampaio, M. E., Alviano, D. S., Axelband, F., Cesar, G. V., Cosentino-Gomes, D., Rodrigues, M. L., Nimrichter, L., Vieyra, A., Alviano, C. S. and Meyer-Fernandes, J. R. (2010). Biochemical characterization of an ecto-ATP diphosphohydrolase activity in Candida parapsilosis and its possible role in adenosine acquisition and pathogenesis. FEMS Yeast Research 10, 735–746.CrossRefGoogle ScholarPubMed
la Sala, A., Ferrari, D., Di Virgilio, F., Idzko, M., Norgauer, J. and Girolomoni, G. (2003). Alerting and tuning the immune response by extracellular nucleotides. Journal of Leukocyte Biology 73, 339–343.CrossRefGoogle ScholarPubMed
Linnertz, H., Thonges, D. and Schoner, W. (1995). Na+/K(+)-ATPase with a blocked E1ATP site still allows backdoor phosphorylation of the E2ATP site. European Journal of Biochemistry 232, 420–424.CrossRefGoogle ScholarPubMed
Menna Barreto, R. F. S., Corrêa, J. R., Pinto, A. V., Soares, M. J. and de Castro, S. L. (2007). Mitochondrial disruption and DNA fragmentation in Trypanosoma cruzi induced by naphthoimidazoles synthesized from beta-lapachone. Parasitology Research 101, 895–905.CrossRefGoogle ScholarPubMed
Meyer-Fernandes, J. R. (2002). Ecto-ATPases in protozoa parasites: looking for a function. Parasitology International 51, 299–303.CrossRefGoogle Scholar
Mizumoto, N., Kumamoto, T., Robson, S. C., Sévigny, J., Matsue, H., Enjyoji, K. and Takashima, A. (2002). CD39 is the dominant Langerhans cell-associated ecto-NTPDase: modulatory roles in inflammation and immune responsiveness. Nature Medicine 8, 358–365.CrossRefGoogle ScholarPubMed
Moreira, O. C., Rios, P. F. and Barrabin, H. (2005). Inhibition of plasma membrane Ca(2+)-ATPase by CrATP. LaATP but not CrATP stabilizes the Ca(2+)-occluded state. Biochimica et Biophysica Acta 1708, 411–419.CrossRefGoogle Scholar
Moreira, O. C., Rios, P. F., Esteves, F. F., Meyer-Fernandes, J. R. and Barrabin, H. (2009). CrATP as a new inhibitor of ecto-ATPases of trypanosomatids. Parasitology 136, 5–44.CrossRefGoogle ScholarPubMed
Mosmann, T. (1983). Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. Journal of Immunological Methods 65, 55–63.CrossRefGoogle ScholarPubMed
Nakaar, V., Beckers, C. J., Polotsky, V. and Joiner, K. A. (1998). Basis for substrate specificity of the Toxoplasma gondii nucleoside triphosphate hydrolase. Molecular and Biochemical Parasitology 97, 209–220.CrossRefGoogle ScholarPubMed
Pauls, H., BredenBröcker, B. and Schoner, W. (1980). Inactivation of (Na+, K+)-ATPase by Chromium (III) complexes of nucleotide triphosphate. European Journal of Biochemistry 109, 523–533.CrossRefGoogle Scholar
Peres-Sampaio, C. E., de Almeida-Amaral, E. E., Giarola, N. L. and Meyer-Fernandes, J. R. (2008). Leishmania amazonensis: effects of heat shock on ecto-ATPase activity. Experimental Parasitology 119, 135–143.CrossRefGoogle ScholarPubMed
Plesner, L. (1995). Ecto-ATPases: identities and functions. International Review of Cytology 158, 141–142.CrossRefGoogle ScholarPubMed
Pinheiro, C. M., Martins-Duarte, E. S., Ferraro, R. B., Fonseca De Souza, A. L., Gomes, M. T., Lopes, A. H., Vannier-Santos, M. A., Santos, A. L. 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
Sansom, F. M., Riedmaier, P., Newton, H. J., Dunstone, M. A., Müller, 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. Journal of Biological Chemistry 283, 12909–12918.CrossRefGoogle ScholarPubMed
Santos, R. F., Pôssa, M. A., Bastos, M. S., Guedes, P. M., Almeida, M. R., Demarco, R., Verjovsky-Almeida, S., Bahia, M. T. and Fietto, J. L. (2009). Influence of Ecto-Nucleoside Triphosphate Diphosphohydrolase Activity on Trypanosoma cruzi Infectivity and Virulence. PloS Neglected Tropical Diseases 3:e387.CrossRefGoogle ScholarPubMed
Souza, V. L., Veras, P. S., Welby-Borges, M., Silva, T. M., Leite, B. R., Ferraro, R. B., Meyer-Fernandes, J. R., Barral, A., Costa, J. M. and Freitas, L. A. (2011). Immune and inflammatory responses to Leishmania amazonensis isolated from different clinical forms of human leishmaniasis in CBA mice. Memorias Instituto Oswaldo Cruz 106, 23–31.CrossRefGoogle ScholarPubMed
Vilsen, B. and Andersen, J. P. (1992 a). CrATP-induced Ca2+ occlusion in mutants of the Ca2+-ATPase of sarcoplasmic reticulum. Journal of Biological Chemistry 267, 25739–25743.CrossRefGoogle ScholarPubMed
Vilsen, B. and Andersen, J. P. (1992 b). Interdependence of Ca2+ occlusion sites in the unphosphorylated sarcoplasmic reticulum Ca2+-ATPase Complex with CrATP. Journal of Biological Chemistry 267, 3539–3550.CrossRefGoogle ScholarPubMed
Walseth, T. F. and Johnson, R. A. (1979). The enzymatic preparation of alpha-[32P]nucleosides triphosphates, cyclic [32P]AMP and cyclic [32P]GMP. Biochimica et Biophysica Acta 562, 11–31.CrossRefGoogle Scholar
World Health Organization (2009). Leishmaniasis: magnitude of the problem. World Health Organization. http://www.who.int/leishmaniasis/burden/magnitude/burden_magnitude/en/index.html (July 2009, last date accessed).Google Scholar
Zimmermann, H. (1999). Two novel families of ectonucleotidases: molecular structures, catalytic properties and a search for a function. Trends in Pharmacological Sciences 20, 231–236.CrossRefGoogle Scholar