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Screening of different classes of proteases in microfilarial and adult stages of Setaria cervi

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Abstract

Many of the filarial proteases involved in critical physiological functions are expressed in stage-specific manner and belong to various mechanistic classes. Setaria cervi, a bovine filarial parasite express different classes of proteases. This parasite shows strong antigenic cross-reactivity with human filarial parasites Wuchereria bancrofti and Brugia malayi. Somatic extracts of S. cervi microfilariae (mf) and adult stages as well as their excretory–secretory (ES) products were screened for the presence of different classes of proteases using general (casein, bovine hemoglobin) and class specific substrates. Detergent-soluble extracts of male and female worms were also screened. Significant enzyme activity was detected in ES products both at pH 5.0 and 7.0 with casein. Cathepsin B-like activity was found to be much higher in membrane-bound extract than in the crude-soluble extract. However, it was also found to be actively secreted by both mf and adult worms. Cathepsin D-like activity assayed at pH 3.0 was very low both in somatic extract as well as in ES products. Collagenase activity at neutral pH showed higher levels, both in somatic extract and ES products. Cathepsin L-like activity was detected only in crude-soluble extract but was below detectable limit in ES products. Leucine aminopeptidase activity was significant both in crude-soluble extract and ES products. This study, thus, might be helpful for a better understanding of host–parasite interaction and identification of appropriate virulence factors that may be targeted as vaccine and/or drug targets against lymphatic filariasis.

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References

  • Acosta D, Goni F, Carmona C (1998) Characterization and partial purification of a leucine aminopeptidase from Fasciola hepatica. J Parasitol 84:1–7

    Article  PubMed  CAS  Google Scholar 

  • Allen JE, Daub J, Guiliano D, McDonnell A, Lizotte-Waniewski M, Taylor DW, Blaxter M (2000) Analysis of genes expressed at the infective larval stage validates utility of Litomosoides sigmodontis as a murine model for filarial vaccine development. Infect Immun 68:5454–5458

    Article  PubMed  CAS  Google Scholar 

  • Britton C, Murray L (2002) A cathepsin L protease essential for Caenorhabditis elegans embryogenesis is functionally conserved in parasitic nematodes. Mol Biochem Parasitol 122:21–33

    Article  PubMed  CAS  Google Scholar 

  • Guiliano DB, Hong X, McKerrow JH, Blaxter ML, Oksov Y, Liu J, Ghedin E, Lustigman S (2004) A gene family of cathepsin L-like proteases of filarial nematodes are associated with larval molting and cuticle and eggshell remodeling. Mol Biochem Parasitol 136:227–242

    Article  PubMed  CAS  Google Scholar 

  • Haffner A, Guilavogui AZ, Tischendorf FW, Brattig NW (1998) Onchocerca volvulus: microfilariae secrete elastinolytic and males nonelastinolytic matrix-degrading serine and metalloproteases. Exp Parasitol 90:26–33

    Article  PubMed  CAS  Google Scholar 

  • Hashmi S, Britton C, Liu J, Guiliano DB, Oksov Y, Lustigman S (2002) Cathepsin L is essential for embryogenesis and development of Caenorhabditis elegans. J Biol Chem 277:3477–3486

    Article  PubMed  CAS  Google Scholar 

  • Hong X, Bouvier J, Wong MM, Yamagata GYL, McKerrow JH (1993) Brugia pahangi: identification and characterization of an aminopeptidase associated with larval molting. Exp Parasitol 76:127–133

    Article  PubMed  CAS  Google Scholar 

  • Jin J, Poole CB, Slatko BE, McReynolds LA (1999) Alternative splicing creates sex-specific transcripts and truncated forms of the furin protease in the parasite Dirofilaria immitis. Gene 237:161–175

    Article  PubMed  CAS  Google Scholar 

  • Jolodar A, Miller DJ (1997) Preliminary characterisation of an Onchocerca volvulus aspartic protease. Int J Parasitol 27:1087–1090

    Article  PubMed  CAS  Google Scholar 

  • Jolodar A, Miller DJ (1998) Identification of a novel family of non-lysosomal aspartic proteases in nematodes. Biochim Biophys Acta 1382:13–16

    PubMed  CAS  Google Scholar 

  • Jolodar A, Fischer P, Buttner DW, Miller DJ, Schmetz C, Brattig NW (2004) Onchocerca volvulus: expression and immunolocalization of a nematode Cathepsin D-like lysosomal aspartic protease. Exp Parasitol 107:145–156

    Article  PubMed  CAS  Google Scholar 

  • Kaushal NA, Kaushal DC, Ghatak S (1987) Identification of antigenic proteins of Setaria cervi by immunoblotting technique. Immunol Invest 16:139–149

    Article  PubMed  CAS  Google Scholar 

  • Kramer JM (1997) In: Riddle DL, Blumenthal T, Meyer BJ, Priess JR (eds) C. elegans II. Cold Spring Harbor Laboratory Press, Plainview, NY, pp 471–500

    Google Scholar 

  • Krithika KN, Dabir P, Kulkarni S, Anandharaman V, Reddy MVR (2005) Identification of 38 kDa Brugia malayi microfilarial protease as a vaccine candidate for lymphatic filariasis. Ind J Exp Biol 43:759–768

    CAS  Google Scholar 

  • Lackey A, James ER, Sakanari JA, Resnick SD, Brown M, Bianco AE, McKerrow JH (1989) Extracellular proteases of Onchocerca. Exp Parasitol 68:176–185

    Article  PubMed  CAS  Google Scholar 

  • Lee DL (2002) Cuticle, moulting and exsheathment. In: Lee DL (ed) The biology of nematodes. Taylor and Francis, London, pp 171–209

    Google Scholar 

  • Lizotte-Waniewski M, Tawe W, Guiliano DB, Lu W, Liu J, Wiliams SA, Lustigman S (2000) Identification of potential vaccine and drug target candidates by expressed sequence tag analysis and immunoscreening of Onchocerca volvulus larval cDNA libraries. Infect Immun 68:3491–3501

    Article  PubMed  CAS  Google Scholar 

  • Loukas A, Selzer PM, Maizels RM (1998) Characterisation of Tc-cpl-1, a Cathepsin L-like cysteine protease from Toxocara canis infective larvae. Mol Biochem Parasitol 92:275–298

    Article  PubMed  CAS  Google Scholar 

  • Lustigman S, McKerrow JH, Shah K, Lui J, Huima T, Hough M, Brotman B (1996) Cloning of a cysteine protease required for the molting of Onchocerca volvulus third stage larvae. J Biol Chem 271:30181–30189

    Article  PubMed  CAS  Google Scholar 

  • Lustigman S, Zhang J, Liu J, Oksov Y, Hashmi S (2004) RNA interference targeting Cathepsin L and Z-like cysteine proteases of Onchocerca volvulus confirmed their essential function during L3 molting. Mol Biochem Parasitol 138:165–170

    Article  PubMed  CAS  Google Scholar 

  • Mallet S, Lesage MC (1987) Relationship between exsheathment and enzyme activity (alkaline phosphatase and leucine amino peptidase) during ageing of Trichostrongylus colubriformis infective larvae. Ann Rech Vet 18:275–278

    PubMed  CAS  Google Scholar 

  • Petralanda I, Yarzabal L, Piessens WF (1986) Studies on a filarial antigen with collagenase activity. Mol Biochem Parasitol 19:51–59

    Article  PubMed  CAS  Google Scholar 

  • Pokharel DR, Rai R, Kumar P, Chaturvedi CM, Rathaur S (2006) Tissue localization of collagenase and leucine aminopeptidase in the bovine filarial parasite Setaria cervi. Filaria journal 5:7

    Article  PubMed  CAS  Google Scholar 

  • Poole CB, Jin J, McReynolds LA (2003) Cloning and biochemical characterization of blisterase, a subtilisin-like convertase from the filarial parasite, Onchocerca volvulus. J Biol Chem 278:36183–36190

    Article  PubMed  CAS  Google Scholar 

  • Ray C, Mckerrow JH (1992) Gut-specific and developmental expression of a Caenorhabditis elegans cysteine protease gene. Mol Biochem Parasitol 51:239–249

    Article  PubMed  CAS  Google Scholar 

  • Richer JK, Sakanari JA, Frank GR, Grieve RB (1992) Dirofilaria immitis: proteases produced by third- and fourth-stage larvae. Exp Parasitol 75:213–222

    Article  PubMed  CAS  Google Scholar 

  • Richer JK, Hunt WG, Sakanari JA, Grieve RB (1993) Dirofilaria immitis: Effect of fluoromethyl ketone cysteine protease inhibitors on the third to fourth-stage molt. Exp Parasitol 76:221–231

    Article  PubMed  CAS  Google Scholar 

  • Roggen DR, Raski DJ, Jones NO (1967) Further electron microscopic observations of Xiphinema index. Nematol 13:1–16

    Article  Google Scholar 

  • Sato K, Nagai Y, Suzuki M (1993) Purification and partial characterization of an acid proteinase from Dirofilaria immitis. Mol Biochem Parasitol 58:293–299

    Article  PubMed  CAS  Google Scholar 

  • Sato K, Yamaguchi H, Waki S, Suzuki M, Nagai Y (1995) Dirofilaria immitis: immunohistochemical localization of acid proteinase in the adult worm. Exp Parasitol 81:63–71

    Article  PubMed  CAS  Google Scholar 

  • Sajid M, McKerrow JH (2002) Cysteine proteases of parasitic organisms. Mol Biochem Parasitol 120:1–21

    Article  PubMed  CAS  Google Scholar 

  • Sharma S, Misra S, Rathaur S (1998) Secretory acetyl cholinesterase of Setaria cervi microfilariae and its antigenic cross-reactivity with Wuchereria bancrofti. Trop Med Int Health 3:46–51

    Article  PubMed  CAS  Google Scholar 

  • Singh RN, Rathaur S (2003) Setaria cervi: In vitro released collagenases and their inhibition by Wuchereria bancrofti infected sera. J Helminthol 77:77–81

    Article  PubMed  CAS  Google Scholar 

  • Skuce PJ, Redmond DL, Liddell S, Stewart EM, Newlands GF, Smith WD, Knox DP (1999) Molecular cloning and characterization of gut-derived cysteine proteinases associated with a host protective extract from Haemonchus contortus. Parasitol 119:405–412

    Article  CAS  Google Scholar 

  • Srivastava Y, Gupta S, Pokharel DR, Rathaur S (2004) Setaria cervi collagenase: IgG cleavage and inhibition by Wuchereria bancrofti infected sera. Acta Parasitol 49:253–259

    CAS  Google Scholar 

  • Tamashiro WK, Rao M, Scott AL (1987) Proteolytic cleavage of IgG and other protein substrates by Dirofilaria immitis microfilarial enzymes. J Parasitol 73:149–154

    Article  PubMed  CAS  Google Scholar 

  • Tort J, Brindley PJ, Knox D, Wolfe KH, Dalton JP (1999) Proteinases and associated genes of parasitic helminths. Adv Parasitol 43:161–266

    Article  PubMed  CAS  Google Scholar 

  • Williamson AL, Brindley PJ, Knox DP, Hotez PJ, Loukas A (2003) Digestive proteases of blood-feeding nematodes. Trends Parasitol 19:417–423

    Article  PubMed  Google Scholar 

Download references

Acknowledgement

The authors are grateful to CSIR, New Delhi scheme no. 37 (1189) EMR-II for the financial support. Daya Ram Pokharel is grateful to University Grants Commission, Nepal for providing partial fellowship for this work.

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  1. Daya Ram Pokharel

    Present address: Department of Biochemistry, Universal College of Medical Sciences, Bhairahawa, Nepal

Authors and Affiliations

  1. Department of Biochemistry, Faculty of Science, Banaras Hindu University, Varanasi-221005, Uttar Pradesh, India

    Elesela Srikanth & Sushma Rathaur

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  1. Daya Ram Pokharel
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  2. Elesela Srikanth
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  3. Sushma Rathaur
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Correspondence to Sushma Rathaur.

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Pokharel, D.R., Srikanth, E. & Rathaur, S. Screening of different classes of proteases in microfilarial and adult stages of Setaria cervi . Parasitol Res 104, 1399–1405 (2009). https://doi.org/10.1007/s00436-009-1336-x

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  • DOI: https://doi.org/10.1007/s00436-009-1336-x

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