Abstract
The ecto protein tyrosine phosphatases (PTP) are known to play a crucial role in the pathogenesis and survival of the intracellular parasites. However, their presence and role in filarial parasites is still unknown. We found a significant amount of tyrosine phosphatase activity in the surface antigen fraction extracted from Setaria cervi (S. cervi), a bovine filarial parasite. An antibody designed against the conserved catalytic core of human protein tyrosine phosphatases, PTP1B cross reacted with a 63 kDa band in the surface antigen. We detected a significant amount of PTP activity in the intact S. cervi adult parasites as well as microfilariae in this study for the first time. This PTP may be localized on the surface of the parasite with an exposed active site available for the external substrates. The PTP activity was also inhibited by sodium orthovanadate and phenyl arsine oxide, specific inhibitors of PTP in both the life stages. The Km and Vmax for PTP in the adult parasites and microfilariae were determined to be 2.574 ± 0.14 mM; 206.3 ± 2.75 μM Pi/h/two parasites and 5.510 ± 0.59 mM; 62.27 ± 2.27 μM Pi/h/106 parasites respectively using O-P-L-Tyrosine as substrate. Interestingly, a positive correlation was observed between the inhibition in PTP activity and reduction in the motility/ viability of the parasites when they were subjected to the specific PTP inhibitors (Orthovanadate and Phenyl arsine oxide) for 4 h in the KRB maintenance medium. The activity was also significantly inhibited in the parasites exposed to antifilarial drug/compounds for e.g. Diethylcarbamazine, Acetylsalicylic Acid and SK7, a methyl chalcone. Therefore suggesting a possible role played by PTP in the survival of the parasite, its interaction with the host as well as in the screening of newly synthesized antifilarials/drugs.
Similar content being viewed by others
References
Aguirre-García MM, Escalona-Montaño AR, Bakalara N, Pérez-Torres A, Gutiérrez-Kobeh L, Becker I (2006) Leishmania major: detection of membrane-bound protein tyrosine phosphatase. Parasitol 132:641–9
Denu JM, Tanner KG (1998) Specific and reversible inactivation of protein tyrosine phosphatases by hydrogen peroxide: evidence for a sulfenic acid intermediate and implications for redox regulation. Biochemistry 37:5633–5642
Dos-Santos AL, Dick CF, Alves-Bezerra M, Silveira TS, Paes LS, Gondim KC, Meyer-Fernandes JR (2012) Interaction between Trypanosoma rangeli and the Rhodnius prolixus salivary gland depends on the phosphotyrosine ecto-phosphatase activity of the parasite. Int J Parasitol 42:819–827
Freire AC, Aoyama H, Haun M, Ferreira CV (2003) Relationship between phosphatase activity and cytotoxic effect of two protein phosphatase inhibitors, okadaic acid and pervanadate, on human myeloid leukemia cell line. J Enzyme Inhib Med Chem 18(5):425–9
Gomes MT, Lopes AH, Meyer-Fernandes JR (2011) Possible roles of ectophosphatases in host-parasite interactions. J Parasitol Res 2011:1–7
Harrington RJ, Gutch MJ, Hengartner MO, Tonks NK, Chisholm AD (2002) The C. elegans LAR-like receptor tyrosine phosphatase PTP-3 and the VAB-1 Eph receptor tyrosine kinase have partly redundant functions in morphogenesis. Development 129:2141–53
Heneberg P (2012) Finding the smoking gun: protein tyrosine phosphatases as tools and targets of unicellular microorganisms and viruses. Curr Med Chem 19:1530–66
Huyer G, Liu S, Kelly J, Moffat J, Pavette P, Kennedy B, Tsaprailis G, Gresser MJ, Ramachandran C (1997) Mechanism of inhibition of protein-tyrosine phosphatases by vanadate and pervanadate. J Biol Chem 272:843–51
Kramer JM (1997) Extracellular matrix in C. elegans II. In: Riddle DL et al (eds) Cold Spring Harbor Laboratory Press. Cold Spring Harbor, New York, pp 471–500
Lemos AP, Pinheiro AAS, Berrêdo-Pinho M, de Souza AF, Motta MCM, De Souza W, Meyer-Fernandes JR (2002) Ectonucleotide diphosphohydrolase activity in Crithidia deanei. Parasitol Res 88:905–911
Lunde ML, Paranjape R, Lawley TJ, Ottesen EA (1988) Filarial antigen in circulating immune complexes from patients with Wuchereria bancrofti filariasis. Am J Trop Med Hyg 38:366–371
Maizels RM, Yazdanbakhsh M (2003) Immune regulation by helminth parasites: cellular and molecular mechanisms. Nat Rev Immunol 3:733–44
Maki J, Yanagisawa T (1980a) Acid phosphatase activity demonstrated in the nematodes, Dirofilaria immitis and Angiostrongylus cantonensis with special reference to the characters and distribution. Parasitol 80:23–38
Maki J, Yanagisawa T (1980b) Comparison of the sites of acid phosphatase activity in an adult filaria. Setaria spp. and in some gastrointestinal nematodes. Parasitol 81:603–608
Martínez-Grueiro MM (2002) Acid phosphatase activity in excretion/secretion products from Heligmosomoides polygyrus adults: an indicator of the physiological status of the worms. Parasitol Res 88:946–9
Meyer-Fernandes JR, Silva-Neto MAC, Soares MS, Fernandes E, Vercesi AE, Oliveira MM (1999) Ecto-phosphatase activities on the cell surface of the amastigote forms of Trypanosoma cruzi. Z Naturforsch C 54(11):977–84
Philipp A, Gómez-Priego MA, Parkhouse RME, Davies MW, Clark NWT, Ogilvie BM, Beltrán-Hernández F (1984) Identification of an antigen of Onchocerca volvulus of possible diagnostic use. Parasitol 89:295–309
Pokharel DR, Srikanth E, Rathaur S (2009) Screening of different classes of proteases in microfilarial and adult stages of Setaria cervi. Parasitol Res 104:1399–405
Rathaur S, Rai R, Srikanth E, Srivastava S (2009) S. cervi dual specific phosphatase, characterization and its effect on eosinophil degranulation. Parasitol 136:895–904
Rathaur S, Yadav M, Singh N, Singh A (2011) Effect of diethylcarbamazine, butylated hydroxy anisole and methyl substituted chalcone on filarial parasite Setaria cervi, proteomic and biochemical approaches. J Proteomics 74:1595–1606
Remaley AT, Das S, Campbell PI, LaRocca GM, Pope MT, Glew RH (1985) Characterization of Leishmania donovani acid phosphatases. J Biol Chem 260:880–6
Ruiz MA, Pérez-Santos JLM, Talamás- Rohana P (2003) An ecto-protein tyrosine phosphatase of Entamoeba histolytica induces cellular detachment by disruption of actin filaments in HeLa cells. Int J Parasitol 33:663–670
Singh A, Rathaur S (2010) Combination of DEC plus aspirin induced mitochondrial mediated apoptosis in filarial parasite Setaria cervi. Biochimie 92:894–900
Stoker AW (2005) Protein tyrosine phosphatases and signalling. J Endocrinol 185:19–33
Wiese M, Berger O, Stierhof YD, Wolfram M, Fuchs MP (1996) Gene cloning and cellular localization of a membrane-bound acid phosphatase of Leishmania mexicana. Mol Biochem Parasitol 82:153–165
World Health Organization (2010) Progress report 2000-2009 and strategic plan 2010-2020 of the global programme to eliminate lymphatic filariasis: halfway towards eliminating lymphatic filariasis. World Health Organization, Geneva
Acknowledgments
The study was supported by CSIR, India (junior and senior research fellowships awarded to NS), Council of Science and Technology UP, India, and Alexander von Humboldt foundation, Germany.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Singh, N., Heneberg, P. & Rathaur, S. Presence of ecto-protein tyrosine phosphatase activity is vital for survival of Setaria cervi, a bovine filarial parasite. Parasitol Res 113, 3581–3589 (2014). https://doi.org/10.1007/s00436-014-4023-5
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00436-014-4023-5