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Classes and crossreactivity of proteinases in the excretory–secretory products of Caenorhabditis elegans

Published online by Cambridge University Press:  01 March 2007

S. Nic an Ultaigh  and
M.F. Ryan
S. Nic an Ultaigh
Affiliation:
School of Biology and Environmental Science, University College Dublin, Belfield, Dublin 4, Ireland
M.F. Ryan*
Affiliation:
School of Biology and Environmental Science, University College Dublin, Belfield, Dublin 4, Ireland
*
*Author for correspondence Fax: 353.1.716.1152 E-mail: MFRyan@ucd.ie
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Abstract

Proteinases released during the in vitro maintenance of asynchronous cultures of the free-living nematode Caenorhabditis elegans were characterized on the basis of subunit composition, fluorogenic substrate specificity, inhibitor sensitivity and pH optima. Cysteine proteinases are present in the excretory–secretory products (ESP) as indicated by the hydrolysis of cathepsin fluorogenic substrates and confirmed by immunoblotting. Serine proteinases were predominant as indicated by substrate gel analysis and inhibitor studies. The presence of metallo-proteinases was also indicated by inhibitor studies. The optimal pH value for cysteine proteinases was 5.5, while serine proteinases were optimal at pH 8.0. As a control, cultures of Escherichia coli, the diet of C. elegans, were extracted separately and gave no evidence of overlap with C. elegans ESP. Cross reactivity between the ESP of C. elegans and antibodies raised against the ESP of the equine parasite Strongylus vulgaris indicated antigenic relatedness of a proteic epitope. This is the first study to characterize the ESP of C. elegans and to display its relatedness with that of S. vulgaris.

Type
Research Papers
Information
Journal of Helminthology , Volume 81 , Issue 1 , March 2007 , pp. 93 - 99
Copyright
Copyright © 2007 Cambridge University Press 2007

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References

Barrett, A.J. (1994) Proteolytic enzymes: serine and cysteine peptidases. Methods in Enzymology 244, 7–11.Google Scholar
Britton, C. & Murray, L. (2002) A cathepsin L protease essential for Caenorhabditis elegans embryogenesis is functionally conserved in parasitic nematodes. Molecular and Biochemical Parasitology 122, 21–33.CrossRefGoogle ScholarPubMed
Britton, C. & Murray, L. (2004) Cathepsin L protease (CPL-1) is essential for yolk processing during embryogenesis in Caenorhabditis elegans. Journal of Cell Science 117, 5133–5143.CrossRefGoogle ScholarPubMed
Britton, C., McKerrow, J.H. & Johnstone, I.L. (1998) Regulation of the Caenorhabditis elegans gut cysteine protease gene cpr-1: requirement for GATA motifs. Journal of Molecular Biology 283, 15–27.CrossRefGoogle ScholarPubMed
Caffrey, C.R. & Ryan, M.F. (1994) Characterization of proteolytic activity of excretory–secretory products from adult Strongylus vulgaris. Veterinary Parasitology 52, 285–296.CrossRefGoogle ScholarPubMed
Cox, G.N., Pratt, D., Hageman, R. & Boisvenue, R.J. (1990) Molecular cloning and primary sequences of a cysteine protease expressed by Haemonchus contortus adult worms. Molecular and Biochemical Parasitology 41, 25–34.CrossRefGoogle ScholarPubMed
Davis, M.W., Birnie, A.J., Chan, A.C., Page, A.P. & Jorgensen, E.M. (2004) A conserved metalloprotease mediates ecdysis in Caenorhabditis elegans. Development 131, 6001–6008.CrossRefGoogle ScholarPubMed
Distefano, S., Palma, J.M., Gomez, M. & Del Rio, L.A. (1997) Characterization of endoproteases from plant peroxisomes. Biochemical Journal 327, 399–405.CrossRefGoogle ScholarPubMed
Dowd, A.J., Dalton, J.P., Loukas, A.C., Prociv, P. & Brindley, P.J. (1994) Secretion of cysteine proteinase activity by the zoonotic hookworm Ancylostoma caninum. American Journal of Tropical Medicine and Hygiene 51, 341–347.CrossRefGoogle ScholarPubMed
Drake, L.J., Bianco, A.E., Bundy, D.A.P. & Ashall, F. (1994) Characterization of peptidases of adult Trichuris muris. Parasitology 109, 623–630.CrossRefGoogle ScholarPubMed
Epstein, H.F. & Shakes, D.C. (1995) Caenorhabditis elegans: modern biological analysis of an organism. Methods in Cell Biology 48, 16–22.Google Scholar
Geldhof, P., Claerebout, E., Knox, D.P., Agneessens, J. & Vercruysse, J. (2000) Proteinases released in vitro by the parasitic stages of the bovine abomasal nematode Ostertagia ostertagi. Parasitology 121, 639–647.CrossRefGoogle ScholarPubMed
Guiliano, D.B., Hong, X., McKerrow, J.H., Blaxter, M., 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. Molecular and Biochemical Parasitology 136, 227–242.CrossRefGoogle ScholarPubMed
Hartmann, S. & Lucius, R. (2003) Modulation of host immune responses by nematode cystatins. International Journal for Parasitology 33, 1291–1302.CrossRefGoogle ScholarPubMed
Hashmi, S., Zhang, J., Oksov, Y. & Lustigman, S. (2004) The Caenorhabditis elegans cathepsin Z-like protease, Ce-CPZ-1, has a multifunctional role during the worm's development. Journal of Biological Chemistry 279, 6034–6045.CrossRefGoogle Scholar
Healer, J., Ashall, F. & Maizels, R.M. (1991) Characterization of proteolytic enzymes from larval and adult Nippostrongylus brasiliensis. Parasitology 103, 305–314.CrossRefGoogle ScholarPubMed
James, G.T. (1978) Inactivation of the protease inhibitor phenylmethylsulfonyl fluoride in buffers. Analytical Biochemistry 86, 574–579.CrossRefGoogle ScholarPubMed
Jasmer, D.P., Roth, J. & Myler, P.J. (2001) Cathepsin B-like cysteine proteases and Caenorhabditis elegans homologues dominate gene products expressed in adult Haemonchus contortus intestine. Molecular and Biochemical Parasitology 116, 159–169.CrossRefGoogle ScholarPubMed
Knox, D.P. & Jones, D.G. (1990) Studies on the presence and release of proteolytic enzymes (proteinases) in gastro-intestinal nematodes of ruminants. International Journal for Parasitology 20, 243–249.CrossRefGoogle ScholarPubMed
Knox, D.P. & Kennedy, M.W. (1988) Proteinases released by the parasitic larval stages of Ascaris suum and their inhibition by antibody. Molecular and Biochemical Parasitology 28, 207–216.CrossRefGoogle ScholarPubMed
Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the heads of bacteriophage T4. Nature 237, 680–685.CrossRefGoogle Scholar
Larminie, C.G.C. & Johnstone, I.L. (1996) Isolation and characterization of four developmentally regulated cathepsin B-like cysteine protease genes from the nematode Caenorhabditis elegans. DNA and Cell Biology 15, 75–82.CrossRefGoogle ScholarPubMed
Loukas, A., Selzer, P.M. & Maizels, R.M. (1998) Characterization of Tc-cpl-1, a cathepsin L-like cysteine protease from Toxocara canis infective larvae. Molecular and Biochemical Parasitology 92, 275–289.CrossRefGoogle ScholarPubMed
MacLennon, K., Gallagher, M.P. & Knox, D.P. (1997) Stage-specific serine and metallo-proteinase release by adult and larval Trichostrongylus vitrinus. International Journal for Parasitology 27, 1031–1036.CrossRefGoogle Scholar
McKerrow, J. (1989) Parasite proteases. Experimental Parasitology 68, 111–115.CrossRefGoogle ScholarPubMed
McKerrow, J. (1999) Development of cysteine protease inhibitors as chemotherapy for parasitic diseases: insights on safety, target validation, and mechanism of action. International Journal for Parasitology 29, 833–837.CrossRefGoogle ScholarPubMed
McKerrow, J.H., Brindley, P., Brown, M., Gam, A., Staunton, C. & Neva, F.A. (1990) Strongyloides stercoralis: identification of a protease that facilitates penetration of skin by the infective larvae. Experimental Parasitology 70, 134–143.CrossRefGoogle ScholarPubMed
Mobarak, M.S. & Ryan, M.F. (1998) An immunohistochemical investigation of the adult stage of the equine parasite Strongylus vulgaris. Journal of Helminthology 72, 159–166.CrossRefGoogle ScholarPubMed
Powers, J.C. & Harper, J.W. (1986) Inhibitors of metalloproteinases. pp. 219–298 in Barrett, A.J. & Salvesen, G. (Eds) Proteinase inhibitors. Amsterdam, Elsevier Science Publishers.Google Scholar
Ray, C. & McKerrow, J.H. (1992) Gut specific and developmental expression of a Caenorhabditis elegans cysteine protease gene. Molecular and Biochemical Parasitology 51, 239–250.CrossRefGoogle ScholarPubMed
Robertson, B.D., Bianco, T., McKerrow, J.H. & Maizels, R.M. (1989) Toxocara canis: proteolytic enzymes secreted by the infective larvae in vitro. Experimental Parasitology 69, 30–36.CrossRefGoogle ScholarPubMed
Sarkis, G.J., Kurpiewski, M.R., Ashcom, J.D., Jen-Jacobson, L. & Jacobson, L. (1988) Proteases of the nematode Caenorhabditis elegans. Archives of Biochemistry and Biophysics 261, 80–90.CrossRefGoogle ScholarPubMed
Silva, F.B., Batista, J.A.N., Marra, B.M., Fragoso, R.R., Monteiro, A.C.S., Figueira, E.L.Z. & Grossi-de-Sa, M.F. (2004) Pro domain peptide of HGCP-Iv cysteine proteinase inhibits nematode cysteine proteinases. Genetics and Molecular Research 3, 342–355.Google ScholarPubMed
Skuce, P.J., Redmond, D.L., Liddell, S., Stewart, E.M., Newlands, G.F.J., Smith, W.D. & Knox, D.P. (1999) Molecular cloning and characterization of gut-derived cysteine proteinases associated with a host protective extract from Haemonchus contortus. Parasitology 119, 405–412.CrossRefGoogle ScholarPubMed
Todorova, V.K., Knox, D.P. & Kennedy, M.W. (1995) Proteinases in the excretory–secretory products (ES) of adult Trichinella spiralis. Parasitology 111, 201–208.CrossRefGoogle ScholarPubMed
Tort, J., Brindley, P.J., Knox, D., Wolfe, K.H. & Dalton, J.P. (1999) Proteinases and associated genes of parasitic helminthes. Advances in Parasitology 43, 161–266.CrossRefGoogle Scholar
Williamson, A.L., Lustigman, S., Oksov, Y., Deumic, V., Plieskatt, J., Mendez, S., Zhan, B., Bottazzi, M.E., Hotez, P. & Loukas, A. (2006) Ancylostoma caninum MTP-1, an astacin-like metalloproteinase secreted by infective hookworm larvae, is involved in tissue migration. Infection and Immunity 74, 961–967.CrossRefGoogle ScholarPubMed